Method for preparation of an extract of at least one bacterium of the species staphylococcus epidermidis, and cosmetic applications of said extract for preventing and / or treating the signs of aging of keratin materials

Abstract

The present invention relates to a method for preparing an extract of at least one bacterium of the species Staphylococcus epidermidis comprising at least one step of culturing at least one bacterium of the species Staphylococcus epidermidis in the presence of a culture medium comprising i) at least one monosaccharide, ii) at least one peptone of vegetable origin and / or at least one yeast extract, and optionally iii) at least one microelement. The present invention is also directed to an extract of at least one bacterium of the species Staphylococcus epidermidis, preferably an extract of at least one bacterial strain of the species Staphylococcus epidermidis selected from the group consisting of bacteria deposited at the CNCM under the serial numbers I-5688, I-5689, I-5904, I-5691, I-5692, I-5693, I-5694 and I-5695, obtainable, obtained or directly obtained by the abovementioned preparation method. The present invention is further directed to a biomass extract of at least one bacterial strain of the species Staphylococcus epidermidis selected from the group consisting of bacteria deposited at the CNCM under the serial numbers I-5688, I-5689, I-5904, I-5691, I-5692, I-5693, I-5694 and I-5695. The present invention also relates to cosmetic treatment methods and cosmetic uses of said extracts for caring for keratin materials, for preventing and / or treating signs of aging of keratin materials, and as active anti-aging agent.

Description

Method for preparation of an extract of at least one bacterium of the speciesStaphylococcus epidermidis, and cosmetic applications of said extract for preventing and / or treating the signs of aging of keratin materials

[0001] The present invention relates to the field of the care of keratin materials, in particular to skin care, more particularly to preventing and / or treating the signs of aging of the skin.

[0002] The present invention relates to a method for preparing an extract of at least one bacterium of the speciesStaphylococcus epidermidiscomprising at least one step of culturing at least one bacterium of the speciesStaphylococcus epidermidisin the presence of a culture medium comprising i) at least one monosaccharide, ii) at least one peptone of vegetable origin and / or at least one yeast extract, and optionally iii) at least one microelement.

[0003] The present invention is also directed to an extract of at least one bacterium of the speciesStaphylococcus epidermidis, preferably an extract of at least one bacterial strain of the speciesStaphylococcus epidermidisselected from the group consisting of bacteria deposited at the CNCM under the serial numbers I-5688, I-5689, I-5904, I-5691, I-5692, I-5693, I-5694 and I-5695, obtainable, obtained or directly obtained by the abovementioned preparation method.

[0004] The present invention is further directed to a biomass extract of at least one bacterial strain of the speciesStaphylococcus epidermidisselected from the group consisting of bacteria deposited at the CNCM under the serial numbers I-5688, I-5689, I-5904, I-5691, I-5692, I-5693, I-5694 and I-5695.

[0005] The present invention also relates to cosmetic treatment methods and cosmetic uses of said extracts for caring for keratin materials, for preventing and / or treating signs of aging of keratin materials, and as active anti-aging agent.Prior art

[0006] Human skin is constituted mainly of two compartments, namely a superficial compartment, the epidermis, and a deep compartment, the dermis, the cohesion of which is ensured by the epidermal junction.

[0007] The epidermis is conventionally divided into a basal layer of keratinocytes that constitutes, as far as the germinal layer of the epidermis, a spinous layer composed of several layers of polyhedral cells positioned on the germinal layers, one to three "granular" layers composed of flat cells containing distinct cytoplasmic inclusions, keratohyalin granules, and finally a set of upper layers referred to as the cornified layers (or stratum corneum), composed of keratinocytes at the terminal stage in their differentiation, referred to as corneocytes.

[0008] The dermis provides the epidermis with a solid support. It is also its nourishing element. It is mainly constituted of fibroblasts and an extracellular matrix.

[0009] Aging of the skin results from various biological processes that can be concomitant: intrinsic aging linked to the passage of time, and extrinsic aging influenced by environmental factors such as exposure to UV or pollution.

[0010] Intrinsic aging or chronological aging is a slow time-dependent process that results in changes in the structure and function of the skin such as the thickness of the skin that is reduced due to atrophy of the dermis and epidermis and in connection with a slowed keratinocyte renewal. Alterations associated with skin aging can manifest themselves in a variety of ways, including:a microrelief characterized in particular by the appearance of fine wrinkles or fine lines, or else by the presence of visible pores in high density, by a roughness altering the surface appearance of the skin;a loss of radiance, resulting in heterogeneity of the complexion;a loss of elasticity, suppleness, firmness, density, tone of the skin, sagging and / or wilting of the skin;a slowing of epidermal renewal leading to a thinning of the stratum corneum of the skin and a papery appearance of the skin.

[0011] In addition, one of the current trends among consumers is that of wanting to minimize the effects of age, to look younger. They therefore seek to attenuate or delay the abovementioned signs of skin aging. Therefore, it is important to identify new active agents to resolve these problems.

[0012] There is therefore a need to identify new active agents that provide one or more anti-aging benefits, in particular new active agents that prevent and / or treat the signs of aging of keratin materials, in particular the skin.

[0013] In particular, there is a need to identify new active anti-aging agents, more particularly new active anti-aging agents making it possible to prevent and / or treat the appearance of a microrelief of the skin and / or fine lines or wrinkles and / or treat a roughness of the skin and / or treat an alteration in the surface appearance of the skin and / or an alteration in the radiance of complexion of the skin, and / or complexion heterogeneity, and / or skin elasticity loss, and / or skin suppleness loss, and / or skin firmness loss, and / or skin density loss, and / or skin tone loss, and / or sagging of the skin, and / or wilting of the skin, and / or thinning of the stratum corneum of the skin and / or a papery appearance of the skin.

[0014] In parallel, the formulation of environmentally friendly cosmetic products, i.e., products whose design and development take account of environmental issues, is becoming a major preoccupation for contributing to meeting the challenges facing the planet. It is therefore proving essential to propose ingredients that make it possible to respond to these environmental issues.

[0015] In this context, it is important to develop new ingredients and / or active agents that are environmentally conscious and are especially of natural origin, in particular biobased, and / or that are derived from sustainable sources that are not obtained from petrochemistry and / or that are biodegradable and / or the process for the extraction of which requires a low consumption of energy and water in order to provide compositions that enable a reduction in the environmental impact of the products.Disclosure of the invention

[0016] The aim of the present invention is to solve the abovementioned technical problems.

[0017] Indeed, the inventors have now discovered that an extract of a bacterium of the speciesStaphylococcus epidermidis,in particular an extract of a bacterium in the form of a lysate of a bacterial strain CNCM-I 5691 of the speciesStaphylococcus epidermidis, makes it possible to increase the thickness of the epidermis, in particular the thickness of thestratum corneum, and therefore has an anti-aging benefit.Summary of the invention

[0018] A first subject of the present invention is a method for preparing an extract of at least one bacterium of the speciesStaphylococcus epidermidiscomprising at least one step of culturing at least one bacterium of the speciesStaphylococcus epidermidisin the presence of a culture medium comprising i) at least one monosaccharide, ii) at least one peptone of vegetable origin and / or at least one yeast extract, and optionally iii) at least one microelement.

[0019] A second subject of the present invention is an extract of at least one bacterium of the speciesStaphylococcus epidermidis, preferably an extract of at least one bacterial strain of the speciesStaphylococcus epidermidisselected from the group consisting of bacteria deposited at the CNCM under the serial numbers I-5688, I-5689, I-5904, I-5691, I-5692, I-5693, I-5694 and I-5695, obtainable, obtained or directly obtained by the abovementioned preparation method.

[0020] A third subject of the present invention is a biomass extract of at least one bacterial strain of the speciesStaphylococcus epidermidisselected from the group consisting of bacteria deposited at the CNCM under the serial numbers I-5688, I-5689, I-5904, I-5691, I-5692, I-5693, I-5694 and I-5695.

[0021] It additionally relates to a composition, in particular cosmetic composition, in particular for caring for keratin materials, comprising, in a physiologically acceptable medium, at least one extract according to the invention.

[0022] A further subject of the present invention is a method for cosmetic treatment of keratin materials, preferably the skin, comprising the application, preferably topically, to the keratin materials, preferably the skin, of a composition according to the invention, for caring for keratin materials, preferably the skin.

[0023] It also relates to the cosmetic use, in particular topical use, of an extract according to the invention, for caring for keratin materials, preferably the skin.

[0024] It also relates to the cosmetic use, in particular topical use, of an extract according to the invention, for preventing and / or treating the signs of aging of keratin materials, preferably for preventing and / or treating the signs of aging of the skin.

[0025] Moreover, another of its subjects is the cosmetic use, in particular topical use, of an extract according to the invention, as an active anti-aging agent.

[0026] Other features, aspects and advantages of the invention will become apparent on reading the detailed description that follows.Definitions

[0027] The term "keratin materials" is understood to mean the skin of the body or face, the lips, the mucous membranes, the eyelashes, the nails, and the hair, of a human being.

[0028] The term “skin” is understood to mean all of the skin of the body and the scalp of a human being, and preferably the skin of the face, neckline, neck, arms and forearms, and hands, or even more preferably still the skin of the face (in particular of the forehead, nose, cheeks, lips and chin), neckline and neck.

[0029] The term “physiologically acceptable medium” is intended to denote a medium that is particularly suitable for applying a composition of the invention to keratin materials, in particular the skin.

[0030] As used herein, the terms “treat” and “treatment” are intended to denote the alleviation of the symptoms associated with a disorder or a specific condition and / or the elimination of said symptoms and also the complete disappearance of the disorder or condition in question.

[0031] In the context of the present invention, the terms “prevent” and “prevention” denote the reduction, to a lesser degree, of the risk or probability of occurrence of a given phenomenon.

[0032] The term "topical use or application" is understood to mean the use or application at the surface of the keratin materials in question, preferably at the surface of the skin in question.

[0033] Staphylococcus epidermidis, or white staphylococcus, is a human commensal bacterium and part of the mucocutaneous flora of virtually the whole of the human population.

[0034] Staphylococcus epidermidis(S. epidermidis) strain CNCM I-5688 (I-5688) is the strain deposited by L’Oréal with the CNCM on June 3, 2021 under accession number CNCM I-5688 (I-5688).

[0035] Staphylococcus epidermidis(S. epidermidis) strain CNCM I-5689 is the strain deposited by L’Oréal with the CNCM on June 3, 2021 under accession number CNCM I-5689 (I-5689).

[0036] Staphylococcus epidermidis(S. epidermidis) strain CNCM I-5904 is the strain deposited by L’Oréal with the CNCM on September 21, 2022 under accession number CNCM I-5904 (I-5904).

[0037] Staphylococcus epidermidis(S. epidermidis) strain CNCM I-5691 is the strain deposited by L’Oréal with the CNCM on June 3, 2021 under accession number CNCM I-5691 (I-5691).

[0038] Staphylococcus epidermidis(S. epidermidis) strain CNCM I-5692 is the strain deposited by L’Oréal with the CNCM on June 3, 2021 under accession number CNCM I-5692 (I-5692).

[0039] Staphylococcus epidermidis(S. epidermidis) strain CNCM I-5693 is the strain deposited by L’Oréal with the CNCM on June 3, 2021 under accession number CNCM I-5693 (I-5693).

[0040] Staphylococcus epidermidis(S. epidermidis) strain CNCM I-5694 is the strain deposited by L’Oréal with the CNCM on June 7, 2021 under accession number CNCM I-5694 (I-5694).

[0041] Staphylococcus epidermidis(S. epidermidis) strain CNCM I-5695 is the strain deposited by L’Oréal with the CNCM on June 7, 2021 under accession number CNCM I-5695 (I-5695).

[0042] Within the meaning of the present invention, the term "extract of at least one bacterium" refers equally to the culture supernatant of said bacteria, the biomass obtained after culturing said bacteria or else the extracts of the biomass that are obtained by treating this biomass.

[0043] The bacterial extract according to the invention may be in a living, semi-active or inactivated, dead, form; the bacterial extract according to the invention is preferably in an inactivated or dead form.

[0044] Advantageously, said extract is distinct from the culture supernatant of said bacteria.

[0045] Advantageously, said extract of at least one bacterium of the speciesStaphylococcus epidermidisis a lysate of at least one bacterium of the speciesStaphylococcus epidermidis.

[0046] Within the meaning of the present invention, a "lysate" means a material obtained after the alteration of bacterial cells, such as the destruction or breakage of bacterial cells, in particular of the cell walls and / or membranes, by a phenomenon known as cell lysis, thus causing the release of intracellular biological constituents naturally contained in the cells of the microorganism in question.

[0047] Within the meaning of the present invention, the term “lysate” is used without prejudice to denote the entirety of the lysate obtained by lysis of the microorganism in question, or only a fraction thereof.

[0048] The lysate used is thus formed totally or partially from the intracellular biological constituents and constituents of the cell walls and membranes.

[0049] More specifically, it contains the cellular cytoplasmic fraction containing enzymes such as lactic acid dehydrogenase, phosphatases, phosphoketolases, transaldolases and metabolites. For illustrative purposes, the constituents of the cell walls are especially peptidoglycan, murein or mucopeptide and teichoic acid, and the constituents of the cell membranes are composed of glycerophospholipids.

[0050] This cell lysis can be accomplished by various technologies well known to those skilled in the art, such as osmotic shock, thermal shock, ultrasound or centrifugation, preferably thermal shock.Detailed description

[0051] Method for preparation of a bacterial extract from the speciesStaphylococcus epidermidis

[0052] A first subject of the present invention is a method for preparing an extract of at least one bacterium of the speciesStaphylococcus epidermidiscomprising at least one step of culturing at least one bacterium of the speciesStaphylococcus epidermidisin the presence of a culture medium comprising i) at least one monosaccharide, ii) at least one peptone of vegetable origin and / or at least one yeast extract, and optionally iii) at least one microelement.

[0053] Preferably, said bacterium of the speciesStaphylococcus epidermidiscultured is a bacterial strain of the speciesStaphylococcus epidermidisselected from the group consisting of bacteria deposited at the CNCM under the serial numbers I-5688, I-5689, I-5904, I-5691, I-5692, I-5693, I-5694 and I-5695, preferably a bacterial strain of the speciesStaphylococcus epidermidisselected from the group consisting of bacteria deposited at the CNCM under the serial numbers I-5688, I-5691, I-5904, I-5692, I-5693, I-5694 and I-5695, even more preferably a bacterial strain of the speciesStaphylococcus epidermidisdeposited at the CNCM under the serial number I-5691.

[0054] Advantageously, said culture medium is free of peptone of animal origin, and preferably said culture medium is free of peptone casein.

[0055] Advantageously, said culture medium comprises less than 4 g / L of sodium chloride, preferably less than 1 g / L by weight of sodium chloride, better still less than 0.5 g / L of sodium chloride, or even is free (0 g / L) of sodium chloride.

[0056] Advantageously, said culture medium is distinct from TSB (tryptic soy broth) medium.

[0057] Within the meaning of the present invention, the term "peptone" is understood to mean a product of a hydrolysis reaction of protein-rich food products. This hydrolysis can be chemical or enzymatic. There are three distinct types of protein raw materials for the manufacture of peptones: peptones of animal origin such as acid casein and whey; peptones of vegetable origin such as soybean, cotton, corn, bean and wheat; and peptones of mycelial origin. Besides the origin of the proteins, peptones can be separated according to their type of hydrolysis: chemical hydrolysis typically by hydrochloric acid, then neutralized by sodium hydroxide; enzymatic hydrolysis, using proteolytic enzymes, digestive enzymes such as pepsin, trypsin and pancreatin, or not, such as papain.

[0058] According to the present invention, the peptone used is a peptone of vegetable origin, preferably a peptone of vegetable origin chosen from soybean, cotton, corn, bean or wheat peptones.

[0059] Advantageously, the peptone used is a peptone obtained by enzymatic hydrolysis, preferably using proteolytic enzymes, more preferably using papain.

[0060] According to one preferred embodiment, the peptone used is a soybean papain peptone.

[0061] Preferably, said culture medium comprises in water, preferably in distilled water:at least one monosaccharide present in the culture medium in an amount ranging from 0.5 to 30 g / L,at least one peptone of vegetable origin present in the culture medium in an amount ranging from 0.5 to 15 g / L, and / or at least one yeast extract present in the culture medium in an amount ranging from 0.5 to 15 g / L, andoptionally at least one microelement present in the culture medium in an amount ranging from 0.05 to 6 mg / L,optionally at least one calcium salt present in the culture medium in an amount ranging from 0.01 to 0.1 g / L.

[0062] Preferentially, said culture medium comprises in water, preferably in distilled water:at least one monosaccharide present in the culture medium in an amount ranging from 0.5 to 7 g / L,at least one peptone of vegetable origin present in the culture medium in an amount ranging from 0.5 to 5 g / L, and / or at least one yeast extract present in the culture medium in an amount ranging from 0.5 to 5 g / L, andoptionally at least one microelement present in the culture medium in an amount ranging from 0.1 to 6 mg / L,optionally at least one calcium salt present in the culture medium in an amount ranging from 0.01 to 0.1 g / L.

[0063] More preferentially, said culture medium comprises in water, preferably in distilled water:at least one monosaccharide present in the culture medium in an amount ranging from 0.5 to 7 g / L, said monosaccharide being chosen from glucose, sucrose, and mixtures thereof,at least one peptone of vegetable origin present in the culture medium in an amount ranging from 0.5 to 5 g / L, said peptone being soybean papain peptone, and at least one yeast extract present in the culture medium in an amount ranging from 0.5 to 5 g / L, said yeast extract being an autolytic yeast extract, andat least one microelement present in the culture medium in an amount ranging from 0.1 to 6 mg / L, the one or more microelements being chosen from Heller microelements, andat least one calcium salt present in the culture medium in an amount ranging from 0.01 to 0.1 g / L, said calcium salt being calcium chloride.

[0064] Even more preferentially, said culture medium comprises in water, preferably in distilled water:at least one monosaccharide present in the culture medium in an amount ranging from 0.5 to 7 g / L, said monosaccharide being glucose,at least one peptone of vegetable origin present in the culture medium in an amount ranging from 0.5 to 5 g / L, said peptone being soybean papain peptone, and at least one yeast extract present in the culture medium in an amount ranging from 0.5 to 5 g / L, said yeast extract being an autolytic yeast extract, andat least one microelement present in the culture medium in an amount ranging from 0.1 to 6 mg / L, the one or more microelements being chosen from Heller microelements, andat least one calcium salt present in the culture medium in an amount ranging from 0.01 to 0.1 g / L, said calcium salt being calcium chloride.

[0065] The Heller microelements, whose composition is given below, were described by Heller, Ann Sci. Nat. Biol. Veg. 14:1 223 (1953). They are mixtures of various mineral elements which were recommended by Heller, not for culturing bacteria, but for the nutrition of plant tissues cultivated in vitro. It should be noted here that no attempt has been made to determine whether the Heller microelements are all indispensable or useful for culturing said bacterium. However, it has been found that the Heller microelements used together in combination with the other abovementioned constituents effectively allow the bacteria in question to be cultured.

[0066] The composition of the Heller microelements, per liter of distilled water, is as follows:

[0067] ZnSO4·7 H2O1 gMnSO4·H2O0.076 gCuSO4·5H2O0.003 gKI0.010 gAlCl3·6H2O0.050 gNiCl2·6H2O0.030 g

[0068] The culturing step may be performed at the appropriate temperature suitable for the bacterial species cultured. Generally, this temperature is between 18 and 40°C. The pH of the culture medium is preferably between 5 and 8.

[0069] When the bacterial culture has reached a terminal stage, that is to say in general when the biomass is at the end of exponential growth, said biomass can be separated from the culture medium. Thus, preferably, the method according to the invention comprises, after the culturing step, a step of separating the bacteria, also called biomass, from the culture medium. This separation step can be obtained by any conventional technique, preferably by centrifugation, filtration, or coagulation, in particular with an alcohol such as ethanol, isopropanol, isobutanol, and mixtures thereof. More preferably, the separation step is a separation of the bacteria (or biomass) from the culture medium by centrifugation.

[0070] The supernatant fraction of this biomass can also be filtered in a sterile container to remove suspended particles, using for example a 0.2 µm filter, and / or stabilized, preferably by heat treatment, by autoclaving for example between 115°C and 121°C for 15 to 40 minutes. This supernatant fraction may also be transferred aseptically into a sterile container. According to one particular embodiment of the invention, the supernatant fraction thus obtained is used as an active anti-aging agent according to the invention.

[0071] Preferably, the method according to the invention comprises, after the separation step, a step of cell lysis. This cell lysis step can be obtained by any conventional technique well known to those skilled in the art, preferably by osmotic shock, thermal shock, ultrasound, or centrifugation, preferentially by thermal shock.

[0072] Preferably, after the step of separating the bacteria (or biomass) from the culture medium, the method according to the invention comprises a step of stabilizing the bacteria (or biomass).

[0073] Preferably, after the cell lysis step, the method according to the invention comprises a step of stabilizing the lysate obtained at the end of the cell lysis step.

[0074] This stabilization step can then be carried out by any method well known to those skilled in the art, preferably by autoclaving, ultra-high temperature (UHT technique), high-pressure sterilization, lyophilization, gamma radiation or freezing, preferentially by autoclaving, even more preferentially by autoclaving carried out at a temperature of between 115°C and 121°C and / or for a time of between 15 and 40 minutes.

[0075] In one preferred embodiment, the stabilization step is carried out by autoclaving carried out at a temperature of between 115°C and 121°C and / or for a time of between 15 and 40 minutes.

[0076] In another preferred embodiment, the stabilization step is carried out by lyophilization.

[0077] A preparation example for an extract which can be used according to the invention is also given in the examples.

[0078] Bacterial extract from the speciesStaphylococcus epidermidis

[0079] The present invention relates to an extract of at least one bacterium of the speciesStaphylococcus epidermidis, preferably an extract of at least one bacterial strain of the speciesStaphylococcus epidermidisselected from the group consisting of bacteria deposited at the CNCM under the serial numbers I-5688, I-5689, I-5904, I-5691, I-5692, I-5693, I-5694 and I-5695, obtainable, obtained or directly obtained by the abovementioned preparation method.

[0080] The extract of at least one bacterium of the speciesStaphylococcus epidermidisaccording to the invention is preferably chosen from i) the culture supernatant of said bacterium, ii) the biomass obtained after culturing said bacterium and iii) the biomass obtained after culturing said bacterium which has undergone a chemical or physical treatment such as a cell lysis treatment and / or a stabilization treatment. Said biomass iii) being preferably in the form of a lysate.

[0081] Preferably, said chemical or physical treatment is chosen from osmotic shock, thermal shock, ultrasound, centrifugation, autoclaving, ultra-high temperature (UHT technique), high-pressure sterilization, lyophilization, gamma radiation, and freezing.

[0082] The present invention also relates to a biomass extract of at least one bacterial strain of the speciesStaphylococcus epidermidisselected from the group consisting of bacteria deposited at the CNCM under the serial numbers I-5688, I-5689, I-5904, I-5691, I-5692, I-5693, I-5694 and I-5695.

[0083] Advantageously, said extract is a lysate of at least one bacterial strain of the speciesStaphylococcus epidermidisselected from the group consisting of bacteria deposited at the CNCM under the serial numbers I-5688, I-5689, I-5904, I-5691, I-5692, I-5693, I-5694 and I-5695.

[0084] Advantageously, said extract is an extract of at least one bacterial strain of the speciesStaphylococcus epidermidisselected from the group consisting of bacteria deposited at the CNCM under the serial numbers I-5688, I-5691, I-5904, I-5692, I-5693, I-5694 and I-5695.

[0085] Advantageously, said extract is a lysate of at least one bacterial strain of the speciesStaphylococcus epidermidisselected from the group consisting of bacteria deposited at the CNCM under the serial numbers I-5688, I-5691, I-5904, I-5692, I-5693, I-5694 and I-5695.

[0086] More preferably, said extract is an extract of at least one bacterial strain of the speciesStaphylococcus epidermidisdeposited at the CNCM under the serial number I-5691.

[0087] Even more preferably, said extract is a lysate of at least one bacterial strain of the speciesStaphylococcus epidermidisdeposited at the CNCM under the serial number I-5691.

[0088] Even better, said extract is a lysate of at least one bacterial strain of the speciesStaphylococcus epidermidisdeposited at the CNCM under the serial number I-5691, obtainable, obtained or directly obtained according to the abovementioned method.

[0089] Advantageously, said extract comprises amino acids; preferably, said amino acids are present in an amount ranging from 20% to 50% by weight relative to the total weight of the dry extract; more preferably, said amino acids are present in an amount ranging from 25% to 45% by weight relative to the total weight of the dry extract.

[0090] The amino acids can be chosen from aspartic acid, threonine, serine, glutamic acid, proline, valine, alanine, cystine, glycine, methionine, isoleucine, leucine, tyrosine, phenylalanine, lysine, arginine, histidine, and mixtures thereof.

[0091] For the purposes of the present invention, the term "amino acids" is understood to mean both free amino acids and those contained in proteins.

[0092] Advantageously, said extract comprises lipids; preferably, said lipids are present in an amount ranging from 1% to 10% by weight relative to the weight of the dry extract; more preferably, said lipids are present in an amount ranging from 3% to 7% by weight relative to the weight of the dry extract.

[0093] Advantageously, said extract comprises sugars; preferably, said sugars are present in an amount ranging from 1% to 10% by weight relative to the weight of the dry extract; more preferably, said sugars are present in an amount ranging from 3% to 7% by weight relative to the weight of the dry extract.

[0094] Advantageously, said extract comprises mineral matter; preferably, said mineral matter is present in an amount ranging from 10% to 30% by weight relative to the weight of the dry extract; more preferably, said mineral matter is present in an amount ranging from 15% to 25% by weight relative to the weight of the dry extract.

[0095] According to one preferred embodiment, said extract comprises:from 20% to 50% by weight of amino acids relative to the total weight of the dry extract, more preferably from 25% to 45% by weight of amino acids relative to the total weight of the dry extract, andfrom 1% to 10% by weight of lipids relative to the weight of the dry extract, more preferably from 3% to 7% by weight of lipids relative to the weight of the dry extract, andfrom 1% to 10% by weight of sugars relative to the weight of the dry extract, more preferably from 3% to 7% by weight of sugars relative to the weight of the dry extract, andfrom 10% to 30% by weight of mineral matter relative to the weight of the dry extract, more preferably from 15% to 25% by weight of mineral matter relative to the weight of the dry extract.Composition according to the invention

[0096] The present invention also relates to a composition, in particular cosmetic composition, in particular for caring for keratin materials, comprising, in a physiologically acceptable medium, at least one extract of at least one bacterial strain of the speciesStaphylococcus epidermidisselected from the group consisting of bacteria deposited at the CNCM under the serial numbers I-5688, I-5689, I-5904, I-5691, I-5692, I-5693, I-5694 and I-5695, obtainable, obtained or directly obtained by the abovementioned preparation method.

[0097] The present invention also relates to a composition, in particular cosmetic composition, in particular for caring for keratin materials, comprising, in a physiologically acceptable medium, at least one extract of at least one bacterial strain of the speciesStaphylococcus epidermidisselected from the group consisting of bacteria deposited at the CNCM under the serial numbers I-5688, I-5689, I-5904, I-5691, I-5692, I-5693, I-5694 and I-5695.

[0098] Advantageously, said extract is present in the composition according to the invention in an amount ranging from 0.0001% to 20% by weight of dry matter relative to the total weight of the composition, preferably in an amount ranging from 0.001% to 10% by weight of dry matter, more preferably in an amount ranging from 0.01% to 5% by weight of dry matter, and better still in an amount ranging from 0.1% to 1% by weight of dry matter, relative to the total weight of the composition.

[0099] Of course, a composition according to the invention may comprise an extract of a single type of bacterial strain as indicated above or a mixture of extracts of at least two or more bacterial strains as listed above.

[0100] A composition according to the invention is preferably devoid of any strain extract different from a strain as listed above.

[0101] The physiologically acceptable medium is generally suited to the nature of the support onto which the composition has to be applied, and also to the appearance under which the composition has to be packaged.

[0102] The physiologically acceptable medium may comprise water and optionally one or more water-miscible solvents.

[0103] According to a preferred embodiment, the compositions of the invention comprise at least 20% by weight of water, in particular at least 40% by weight of water relative to the total weight of said composition. Preferentially, the composition according to the invention has a water content ranging from 20% to 95% by weight, even better still from 40% to 90% by weight, relative to the total weight of the composition.

[0104] The water-miscible (at room temperature such as 25°C) organic solvents include lower monoalcohols having from 1 to 5 carbon atoms such as ethanol and isopropanol, glycols having from 2 to 8 carbon atoms such as ethylene glycol, hexylene glycol, propylene glycol, 1,3-butylene glycol, pentylene glycol and dipropylene glycol, and glycerol.

[0105] The water-miscible organic solvent(s) may be present in a concentration of from 0.01% to 30% by weight, preferably from 0.1% to 20% by weight and more preferably from 1% to 15% by weight, relative to the total weight of the composition.

[0106] According to one particular embodiment, the pH of the composition according to the invention is between 4 and 7.5, notably between 4.5 and 7, and in particular between 4.7 and 6.5.

[0107] The composition according to the invention may also comprise any water-soluble or water-dispersible compound such as gelling agents, film-forming polymers, thickeners, surfactants, and mixtures thereof.

[0108] The composition according to the invention may also comprise at least one fatty substance such as one or more oils.

[0109] According to one embodiment, the composition may comprise from 5% to 80% by weight of fatty substance, preferably of oil(s), and preferably from 10% to 60% of fatty substance, preferably of oil(s), by weight relative to the total weight of said composition.

[0110] The term “oil” refers to any fatty substance that is in liquid form at ambient temperature (20-25°C) and at atmospheric pressure. These oils may be of plant, mineral or synthetic origin.

[0111] As oils that can be used in the composition of the invention, examples that may be mentioned include:

[0112] - hydrocarbon oils of plant origin;

[0113] - synthetic esters and ethers, notably of fatty acids, for instance oils of formulae R1COOR2 and R1OR2 in which R1 represents the residue of a fatty acid comprising from 8 to 29 carbon atoms, and R2 represents a branched or unbranched hydrocarbon chain containing from 3 to 30 carbon atoms;

[0114] - linear or branched hydrocarbons, of mineral or synthetic origin;

[0115] - fatty alcohols having from 8 to 26 carbon atoms;

[0116] - fluoro oils which are partially hydrocarbon-based and / or silicon-based;

[0117] - silicone oils;

[0118] - mixtures thereof.

[0119] In the list of oils mentioned above, a hydrocarbon oil means any oil mainly comprising carbon and hydrogen atoms, and optionally ester, ether, fluoro, carboxylic acid and / or alcohol groups.

[0120] According to a preferred embodiment of the invention, the composition used in the context of the invention is a water-in-oil (W / O) or oil-in-water (O / W) emulsion, preferably an oil-in-water (O / W) emulsion. The proportion of fatty substance in the emulsion may range from 5% to 80% by weight and preferably from 10% to 60% by weight relative to the total weight of the composition.

[0121] The composition may comprise at least one emulsifier. The emulsions generally contain at least one emulsifier chosen from amphoteric, anionic, cationic and non-ionic emulsifiers, used alone or as a mixture, and optionally a coemulsifier. The emulsifiers are appropriately chosen according to the emulsion to be obtained (W / O or O / W).

[0122] The emulsifier and the coemulsifier are generally present in the composition in a proportion ranging from 0.3% to 20% by weight, and preferably from 0.5% to 10% by weight, relative to the total weight of the composition.

[0123] Preferably, a composition according to the invention may comprise at least one additional ingredient selected from the group consisting of fatty substances, organic solvents, thickeners, fragrances, preservatives, surfactants, active cosmetic agents other than the extract according to the invention, such as active anti-aging agents other than the extract according to the invention, fillers and mixtures thereof.

[0124] Such an additional ingredient may represent from 0.0001% to 20%, preferably from 0.01% to 10% and better still from 0.1% to 5% by weight, relative to the total weight of the composition.

[0125] Of course, a person skilled in the art will take care to select these one or more additional ingredients, and / or the amount thereof, such that the advantageous properties of the strains according to the invention are not, or not substantially, adversely affected by the envisioned addition.

[0126] A composition according to the invention may be in any presentation form normally used in the cosmetics field.

[0127] It may notably be in the form of an aqueous or aqueous-alcoholic solution, which may be gelled, a dispersion of the lotion type, which may be a two-phase dispersion, an oil-in-water or water-in-oil emulsion or a multiple emulsion, an aqueous gel, or else a dispersion of oils in an aqueous phase, notably using spherules, it being possible for these spherules to be polymeric particles or, better still, lipid vesicles of ionic and / or nonionic type. It may be of more or less fluid liquid consistency.

[0128] Preferentially, a composition according to the invention differs from compositions having an essentially detergent purpose with regard to the skin, hair and / or mucous membranes, such as soaps, shampoos and shower gels for washing and / or cleansing.

[0129] A composition according to the invention is preferentially suitable for topical administration.

[0130] Thus, a composition according to the invention may comprise all the constituents usually employed in the envisaged topical application and administration.

[0131] A composition according to the invention may advantageously be in the form of an emulsion, particularly obtained by dispersion of an aqueous phase in a fatty phase (W / O) or of a fatty phase in an aqueous phase (O / W), of liquid or semi-liquid consistency of the milk type, or of soft consistency, or else of a multiple emulsion (W / O / W or O / W / O). These compositions are prepared according to the usual known methods.

[0132] More particularly, a composition according to the invention may be intended for topical application and may preferably be in the form of an emulsion, preferably an oil-in-water emulsion. Preferably, such an emulsion is not intended to be rinsed off after application.

[0133] A composition according to the invention is preferentially intended to be applied to keratin materials, preferably the skin.

[0134] A composition alternatively can be in the form of a face and / or body care or makeup product, and can be packaged, for example, in the form of a cream in a jar or a fluid in a tube or a pump bottle or a dropper bottle.

[0135] A composition according to the invention can be manufactured by any known process generally used in the cosmetics field.

[0136] The ingredients are mixed before shaping, in the order and under conditions readily determined by a person skilled in the art.

[0137] According to a particular mode of the invention, other agents intended to embellish the appearance and / or texture of the skin may also be added to a composition according to the invention.Uses and processes

[0138] According to one of its aspects, the present invention relates to a method for cosmetic treatment of keratin materials, preferably the skin, comprising the application, preferably topically, to the keratin materials, preferably the skin, of a composition according to the invention, for caring for keratin materials, preferably the skin. More preferably, the cosmetic treatment method according to the invention is intended for the prevention and / or treatment of the signs of aging of the skin.

[0139] The present invention also relates to the cosmetic use, in particular topical use, of an extract according to the invention, for caring for keratin materials, preferably the skin.

[0140] The present invention further relates to the cosmetic use, in particular topical use, of an extract according to the invention, for preventing and / or treating the signs of aging of keratin materials, preferably for preventing and / or treating the signs of aging of the skin.

[0141] Moreover, another of its subjects is the cosmetic use, in particular topical use, of an extract according to the invention, as an active anti-aging agent.

[0142] The term "signs of skin aging" is understood to mean any modifications of the external appearance of the skin due to aging.

[0143] The signs of skin aging targeted by the invention may be chosen from an appearance of a microrelief of the skin, a formation and / or a presence of fine lines or wrinkles, an appearance of roughness, an alteration of the surface appearance of the skin, altered radiance of complexion of the skin, heterogeneity of the complexion, loss of elasticity, loss of suppleness, loss of firmness, loss of density, or loss of tone of the skin, sagging and / or wilting of the skin, a thinning of the stratum corneum of the skin, and a papery appearance of the skin.

[0144] Preferably, the signs of skin aging targeted by the invention are chosen from the appearance of a microrelief of the skin, the formation and / or the presence of fine lines or wrinkles, the appearance of roughness, the alteration of the surface appearance of the skin, thinning of the stratum corneum of the skin, and the papery appearance of the skin.

[0145] More preferably, the signs of skin aging targeted by the invention are chosen from the appearance of a microrelief of the skin, the formation and / or the presence of fine lines and / or wrinkles, the appearance of roughness, the alteration of the surface appearance of the skin, and thinning of the stratum corneum of the skin.

[0146] Even better, the sign of skin aging targeted by the invention is the thinning of the stratum corneum of the skin.

[0147] The skin is as defined previously. Preferably, the skin is aged. The term "aged skin" means a general esthetic state of the skin resulting from aging.

[0148] The cosmetic uses and methods considered according to the invention are non-therapeutic.

[0149] The cosmetic uses and methods of the invention are preferentially performed by topically administering an extract according to the invention or a composition according to the invention.

[0150] By way of illustration, the cosmetic use or method according to the invention may be implemented by topical application, daily for example, of at least one composition according to the invention, which may be formulated, for example, in the form of a cream, gel, serum, lotion, emulsion, or makeup-removing milk, preferably in the form of an emulsion.

[0151] Application may be repeated, for example, 1 to 2 times daily for one or more days and generally over an extended period of at least 4 weeks, or even 4 to 15 weeks, with, where appropriate, one or more periods of stoppage.

[0152] According to one embodiment, the application is daily (once a day) and generally over a prolonged period of at least 4 weeks, or even 4 to 15 weeks, with, where appropriate, one or more periods of stoppage.

[0153] According to one embodiment, the cosmetic treatment method according to the invention may comprise a single application.

[0154] Throughout the description, including the claims, the terms “between ... and ...”, and “ranging from ... to ...” should be understood as meaning limits included, unless otherwise specified.

[0155] The examples that follow illustrate the present invention without limiting the scope thereof.

[0156] In the examples, unless otherwise specified, the temperature is room temperature (20°C) and is expressed in degrees Celsius, and the pressure is atmospheric pressure.Examples

[0157] Example 1: Preparation of an extract according to the invention:

[0158] A culture of theStaphylococcus epidermidisstrain CNCM-I 5691 is carried out in its complete culture medium in a 10 effective liter bioreactor in batch or fed batch mode. During this step, the pH is regulated at 6.5 by adding a 1M KOH solution, the temperature is maintained at 32°C and the dissolved oxygen at 40%.

[0159] The composition of the initial culture medium is described in Table 2 below.

[0160] Chemical namesConcentrations by massAutolytic yeast extract5 g / LSoybean papain peptone5 g / LGlucose6.5 g / LKH2PO40.088 g / LCaCl20.050 g / LCuSO4.5H2O60 µg / LMnSO4.1H2O152 µg / LKI20 µg / LZnSO4.7H2O200 µg / LAlCl3.6H2O100 µg / Ldeionized waterqs for 1 L

[0161] As soon as the plateau phase is reached after 20 to 48 hours of culture, the extraction and the separation of the cells are carried out by centrifugation (under laboratory conditions, 10 000 g for 30 min). Then the pellet, also called biomass, containing the cells is recovered, to be subsequently frozen at -20°C and then thawed, allowing the cells to burst and thus a lysate to be obtained. The lysate is then packaged in bags or tubes and finally stabilized by sterilization at 121°C for 30 minutes.

[0162] The lysate obtained at the end of the method as described according to the example contains 5.2% by weight of dry matter, relative to the total weight of the lysate.

[0163] The analysis of the constituents of the dry extract is presented in Table 3 below.

[0164] Compositiong per 100 g of dry extractTotal lipids5.88Total amino acids34.30Total sugars5.29Mineral matter19.01Example 2: Modulation of epidermal thickness

[0165] The RHE epidermal thickness modulation (RH3E) evaluation test is carried out to evaluate the efficacy of raw materials, including the extract ofStaphylococcus epidermidisobtained according to example 1, on the 3D in vitro SkinEthic RHE® model after systemic application, for 4 or 7 days.Biological model

[0166] The effects of the raw materials tested, including the extract ofStaphylococcus epidermidisobtained according to example 1, are determined by an evaluation of the quality of the reconstructed epidermis as a function of its thickness and morphology. The experiments are carried out on the SkinEthic RHE® reconstructed epidermal model.

[0167] The tissues are received on medium on D10 and stored at 37°C (95% humidity and 5% carbon dioxide) until the end of the treatments on D14 or D17. Systemic treatments are carried out on D10, D12 and stopped on D14 (short-term protocol) or stopped on D17 (long-term protocol). The tissues are cultured with 2 mL of treatment solution (raw material (RM) in SGM medium) in a 6-well plate and stored at 37°C (95% humidity and 5% carbon dioxide) between treatments.Treatments

[0168] For all the conditions tested, the raw materials are tested in biological triplicate (n=3). For the raw materials, if no toxicity is observed on D14 and good histological quality is noted, it is possible to treat the tissues on D14 with 3 mL of treatment solution and harvest them on D17 (long-term protocol).OCT acquisition and analyses

[0169] OCT (optical coherence tomography) is a non-invasive imaging technique based on the analysis of infrared light reflected from tissues and the creation of an interference signal. OCT allows visualization of surface appearance; the 2D structure of the RHE® model is followed during epidermal reconstruction. This also makes it possible to determine, by acquisition and segmentation using algorithms specifically developed by Newtone®, the epidermal thickness of the tissue after reconstruction.

[0170] Image acquisition and analysis are performed on D10, D12, D14 and D17 (if a longer protocol is performed) over all the conditions tested.Active agents tested

[0171] For each molecule tested, the interpretation criteria are based on:Modulation of epidermal thickness,Histological quality at the end of treatment.

[0172] Each condition is compared to the vehicle (untreated control or DMSO control).RESULTS FOR BIOLOGICAL ACTIVITY

[0173] The extract ofStaphylococcus epidermidisobtained according to example 1 enhances epidermal quality after systemic application, as shown in Table 4 below.

[0174] The extract ofStaphylococcus epidermidisobtained according to example 1 was tested in systemic application on a SkinEthic® RHE model for 4 days and 7 days and at several applications. Several parameters were measured.

[0175] The application of the extract ofStaphylococcus epidermidisobtained according to example 1:Slightly increases (p-value=0.017) epidermal thickness at 7 days and at a dose of 250 µg / ml: +8.9% variation (+ / - 3.12),Significantly increases (p-value=0.000) the stratum corneum layer: +20.6% (+ / - 1.7) at a dose of 250 µg / ml.

[0176] Starting materials testedOverall thicknessVariation in epidermal thickness on D7Variation in thickness of the stratum corneum on D7µM% variationStandard deviationp-value% variationStandard deviationp-valueVehicle1060.6%3.120.8392%1.70.912Retinol12630.1%3.120.0007.2%1.70.000Staphylococcus epidermidisextract obtained according to example 11128.97%3.120.01720.6%1.70.000

Claims

A method for preparing an extract of at least one bacterium of the speciesStaphylococcus epidermidiscomprising at least one step of culturing at least one bacterium of the speciesStaphylococcus epidermidisin the presence of a culture medium comprising i) at least one monosaccharide, ii) at least one peptone of vegetable origin and / or at least one yeast extract, and optionally iii) at least one microelement.The preparation method as claimed in claim 1, wherein said culture medium comprises in water:at least one monosaccharide present in the culture medium in an amount ranging from 0.5 to 30 g / L, andat least one peptone of vegetable origin present in the culture medium in an amount ranging from 0.5 to 15 g / L, and / or at least one yeast extract present in the culture medium in an amount ranging from 0.5 to 15 g / L, andoptionally at least one microelement present in the culture medium in an amount ranging from 0.05 to 6 mg / L, andoptionally at least one calcium salt present in the culture medium in an amount ranging from 0.01 to 0.1 g / L.The preparation method as claimed in any one of the preceding claims, wherein said bacterium of the speciesStaphylococcus epidermidiscultured is a bacterial strain of the speciesStaphylococcus epidermidisselected from the group consisting of bacteria deposited at the CNCM under the serial numbers I-5688, I-5689, I-5904, I-5691, I-5692, I-5693, I-5694 and I-5695, preferably a bacterial strain of the speciesStaphylococcus epidermidisselected from the group consisting of bacteria deposited at the CNCM under the serial numbers I-5688, I-5691, I-5904, I-5692, I-5693, I-5694 and I-5695, even more preferably a bacterial strain of the speciesStaphylococcus epidermidisdeposited at the CNCM under the serial number I-5691.The preparation method as claimed in any one of the preceding claims, characterized in that it comprises, after said culturing step, a step of separating the bacteria from the culture medium, preferably by centrifugation, filtration, or coagulation, more preferably by centrifugation.The preparation method as claimed in the preceding claim, characterized in that it comprises, after said separation step, a step of cell lysis, preferably by osmotic shock, thermal shock, ultrasound, or centrifugation, more preferably by thermal shock.The preparation method as claimed in the preceding claim, characterized in that it comprises, after said cell lysis step, a stabilization step, preferably by autoclaving, ultra-high temperature, high-pressure sterilization, lyophilization, gamma radiation or freezing, preferably by autoclaving.An extract of at least one bacterium of the speciesStaphylococcus epidermidis, preferably of at least one bacterial strain of the speciesStaphylococcus epidermidisselected from the group consisting of bacteria deposited at the CNCM under the serial numbers I-5688, I-5689, I-5904, I-5691, I-5692, I-5693, I-5694 and I-5695, obtainable by the preparation method as defined according to any one of claims 1 to 6.The extract as claimed in claim 7, characterized in that it takes the form of a lysate.A composition, in particular cosmetic composition, in particular for caring for keratin materials, comprising, in a physiologically acceptable medium, at least one extract as defined according to any one of claims 7 to 8.The composition as claimed in claim 9, wherein said extract is present in the composition in an amount ranging from 0.0001% to 20% by weight of dry matter relative to the total weight of the composition, preferably in an amount ranging from 0.001% to 10% by weight of dry matter, more preferably in an amount ranging from 0.01% to 5% by weight of dry matter, and better still in an amount ranging from 0.1% to 1% by weight of dry matter, relative to the total weight of the composition.A method for cosmetic treatment of keratin materials, preferably the skin, comprising the application, preferably topically, to the keratin materials, preferably the skin, of a composition as defined according to any one of claims 9 and 10, for caring for keratin materials, preferably the skin.The cosmetic use, in particular topical use, of an extract as defined according to any one of claims 7 to 8, for caring for keratin materials, preferably the skin.The cosmetic use, in particular topical use, of an extract as defined according to any one of claims 7 to 8, for preventing and / or treating the signs of aging of keratin materials, preferably for preventing and / or treating the signs of aging of the skin.The cosmetic use, in particular topical use, of an extract as defined according to any one of claims 7 to 8, as an active anti-aging agent.A biomass extract of at least one bacterial strain of the speciesStaphylococcus epidermidisselected from the group consisting of bacteria deposited at the CNCM under the serial numbers I-5688, I-5689, I-5904, I-5691, I-5692, I-5693, I-5694 and I-5695.Composition, in particular cosmetic composition, in particular for caring for keratin materials, comprising, in a physiologically acceptable medium, at least one biomass extract as defined according to claim 15.Composition as claimed in claim 16, wherein said biomass extract is present in the composition in an amount ranging from 0.0001% to 20% by weight of dry matter relative to the total weight of the composition, preferably in an amount ranging from 0.001% to 10% by weight of dry matter, more preferably in an amount ranging from 0.01% to 5% by weight of dry matter, and better still in an amount ranging from 0.1% to 1% by weight of dry matter, relative to the total weight of the composition.