COMPOSITION AND PROCESS FOR THE TREATMENT OF KERATINOUS MATERIALS

Abstract

COMPOSITION AND METHOD FOR THE CARE OF KERATINOUS MATERIALS The present invention relates to a composition, preferably for the care of keratinous materials, comprising: a) at least one microorganism of the species of the genus Bifidobacterium, the microorganism being in lysate form, b) at least one extract of Salvia miltiorrhiza, and c) an extract of Sphingomonas ferment. The present invention also relates to a non-therapeutic method for the care of keratinous materials, comprising the application of said composition to the keratinous materials. Figure for the abstract: none

Description

Title of the invention: COMPOSITION AND METHOD FOR THE CARE OF KERATINOUS MATERIALS technical field

[0001] The present invention relates to a cosmetic composition. In particular, the present invention relates to a keratinous material care composition. The present invention also relates to a non-therapeutic method for caring for keratinous materials. CONTEXT OF THE INVENTION

[0002] The skin is the protective barrier of the human body. It protects the body's interior against physical damage (such as trauma) and biological damage (such as that caused by bacteria, viruses, or fungi). The epidermis is a keratinized stratified squamous epithelium. Its average thickness ranges from 60 to 100 µm and can reach 600 to 700 µm on the soles of the feet and the palms of the hands. It is composed mainly of keratinocytes, but also of other cells, and rests on a basement membrane that separates it from the dermis.

[0003] The main changes concern the dermis and are a decrease in collagen content and dermal thickness.

[0004] The main changes concerning the epidermis are a decrease in the differentiation of keratinocytes, resulting in a deficiency of the protein matrix of the corneal cell, an increase in metalloproteinases, which are proteases that degrade the extracellular matrix and participate in skin aging, as well as a decrease in the synthesis of various glycosaminoglycans.

[0005] The development of formulas dedicated to skin care is ongoing.

[0006] A wide variety of cosmetic compositions have been used for skin care, for example to provide a repairing and soothing effect on the skin. However, for most of them, the skin care effect, particularly the repairing and soothing effect, is not satisfactory.

[0007] Thus, there is always a need to formulate a composition to take care of the skin, which can provide a good repairing and / or soothing effect. Summary of the invention

[0008] An object of the present invention is therefore to develop a skincare composition of keratinous materials, which can deliver a good repairing and / or soothing effect.

[0009] Another object of the present invention is to propose a non-therapeutic method for the treatment of keratinous materials.

[0010] Accordingly, in a first aspect, the present invention proposes a composition, preferably for the care of keratinous materials, comprising: a. at least one microorganism of the species of the genus Bifidobacterium, the microorganism being in the form of a lysate, b. at least one extract of Salvia miltiorrhiza, and c. an extract of ferment of Sphingomonas.

[0011] In a second aspect, the present invention proposes a non-therapeutic method for the treatment of keratinous materials, comprising the application of the composition according to the first aspect of the present invention to the keratinous materials.

[0012] The composition of the present invention can provide a good repairing and / or soothing effect to keratinous materials.

[0013] Other subjects, features, aspects, and advantages of the present invention will be presented in the following description and will be partly evident from the description or may be learned through practice of the present invention. DETAILED DESCRIPTION OF THE INVENTION

[0014] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as that commonly understood by a person skilled in the art of the present invention. Where the definition of a term in this description conflicts with the meaning commonly understood by a person skilled in the art of the present invention, the definition described herein shall apply.

[0015] In what follows, and unless otherwise indicated, the limits of a range of values ​​are included in that range, in particular in the expressions "between... and..." and "from... to...".

[0016] Furthermore, the expression "at least one" used in this description is equivalent to the expression "one or more".

[0017] Throughout this application, the term "including" shall be interpreted as encompassing all the specifically cited features as well as any optional, additional, and unspecified features. As used herein, the use of the term "including" also discloses the embodiment in which no features other than the specifically cited features are present (i.e., "consisting of").

[0018] Unless otherwise stated, all numerical values ​​expressing a quantity of ingredients and the like used in the description and claims shall be understood as modified by the term "approximately". Accordingly, unless otherwise stated, the numerical values ​​and parameters described These are approximate values ​​which, if necessary, can be changed according to the desired objective.

[0019] For the purposes of the present invention, the term "keratinous materials" is intended to refer to human skin, in particular facial skin and lips. Facial skin is specifically considered in the context of the present invention.

[0020] In the present invention, all percentages refer, unless otherwise specified, to a percentage by weight.

[0021] According to the first aspect, the composition of the present invention comprises: a. at least one microorganism of the species of the genus Bifidobacterium, the microorganism being in lysate form, b. at least one extract of Salvia miltiorrhiza, and c. an extract of Sphingomonas ferment.

[0022] Microorganism of species of the genus Bifidobacterium

[0023] According to the first aspect, the composition of the present invention comprises at least one microorganism of the species of the genus Bifidobacterium, the microorganism being in the form of a lysate.

[0024] A lysate commonly refers to a substance obtained after the destruction or dissolution of biological cells by a phenomenon called cell lysis, which therefore causes the release of intracellular biological constituents naturally contained in the cells of the microorganism concerned.

[0025] In the context of the present invention, the term "lysate" is used interchangeably to designate the entire lysate obtained by the lysis of the microorganism concerned or only a fraction of the lysate. The lysate used is thus formed in whole or in part from intracellular biological constituents and from constituents of cell walls and membranes.

[0026] More specifically, the lysate contains the cytoplasmic cellular fraction containing enzymes, such as lactic acid dehydrogenase, phosphatases, phosphoketolases or transaldolases, and metabolites. By way of illustration, the constituents of cell walls are in particular peptidoglycan, murein or mucopeptide and teichoic acid, and the constituents of cell membranes are composed of glycerophospholipids.

[0027] This cell lysis can be achieved by various techniques, for example by osmotic shock, thermal shock, ultrasound, or under mechanical stress, for example by centrifugation. More particularly, this lysate can be obtained according to the technology described in US patent 4,464,362 and in particular according to the following protocol.

[0028] The microorganism(s) of the species of the genus Bifidobacterium is / are preferably cultured under anaerobic conditions in a suitable culture medium, For example, according to the conditions described in US documents 4,464,362 and EP 4376804. Once the stationary phase of development is reached, the culture medium can be inactivated by pasteurization at a temperature ranging, for example, from 60 to 65 °C for 30 minutes. The microorganism(s) thus formed are then collected by a conventional separation technique, such as membrane filtration or centrifugation, and resuspended in a sterile physiological saline solution of NaCl.

[0029] The lysate of these microorganisms can then be obtained by ultrasonic disintegration of the solution obtained above, so as to release the cytoplasmic fractions, cell wall fragments, and metabolic products. All components, in their natural distribution, are then stabilized in a weakly acidic aqueous solution. The total content of active substance(s) present in the lysate obtained by this process preferably ranges from 0.1% to 50% by weight, more preferably from 1% to 20% by weight, and even better, this content is 5% by weight, relative to the total weight of the lysate.

[0030] The lysate can be used in different forms, in particular as a solution or in powder form.

[0031] Preferably, the microorganism of species of the genus Bifidobacterium is chosen from the following species: Bifidobacterium longum, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium animalis, Bifidobacterium lactis, Bifidobacterium infantis, Bifidobacterium adolescentis, Bifidobacterium pseudobacterium, and mixtures thereof.

[0032] More preferably, the microorganism of species of the genus Bifidobacterium is Bifidobacterium longum.

[0033] In particular, we can cite, as a usable lysate within the meaning of the invention, the ingredient bearing the INCI name: Bifida Ferment Lysate.

[0034] In particular, one can cite, by way of example, the product sold under the name Repair Complex CLR® by K. Richter GmbH, which is formed from an inactivated lysate of the species Bifidobacterium longum.

[0035] Advantageously, the microorganism of species of the genus Bifidobacterium is present in the composition of the present invention in an amount ranging from 0.001% by weight to 20% by weight, preferably from 0.01% by weight to 15% by weight, more preferably from 0.01% by weight to 10% by weight, relative to the total weight of the composition. Salvia miltiorrhiza extracts

[0036] According to the first aspect, the composition according to the present invention comprises at least one extract of Salvia miltiorrhiza.

[0037] Preferably, the Salvia miltiorrhiza extract is chosen from a Salvia miltiorrhiza root extract and a Salvia miltiorrhiza leaf extract.

[0038] More preferably, the composition of the present invention comprises an extract of Salvia miltiorrhiza root.

[0039] Advantageously, the Salvia miltiorrhiza root extract comprises at least 5% by weight of total polyphenols relative to the total dry weight of the extract, preferably from 5% by weight to 15% by weight of total polyphenols, relative to the total dry weight of the extract.

[0040] In particular, we can cite, as an example, the product sold under the name NEUROFENSE MC by SILAB, which is an extract of Salvia miltiorrhiza root.

[0041] Advantageously, the extract of Salvia miltiorrhiza is present in the composition of the present invention in an amount ranging from 0.1 ppm to 50,000 ppm, preferably from 0.1 ppm to 10,000 ppm, more preferably from 0.5 ppm to 5,000 ppm, and even more preferably from 1 ppm to 2,500 ppm, relative to the total weight of the composition. Sphingomonas ferment extract

[0042] According to the first aspect, the composition of the present invention comprises a ferment extract of Sphingomonas.

[0043] Sphingomonas ferment extract is the extract of the product obtained by the fermentation of Sphingomonas. It is known to contain glycosphingolipids (GSLs) such as seramide, dihydrosphingosine, and glucuronic acid, and is valued for its reduction of lipid peroxidation and its effect on key biomarkers of senescence, thereby preventing skin aging.

[0044] The extracts according to the invention can be prepared by any conventional process well known to those skilled in the art. These processes generally include a step of fermentation, isolation, separation and purification.

[0045] Bacteria of the genus Sphingomonas are generally cultured in a suitable medium and grown to a desired density. Preferably, the bacteria themselves are concentrated by any known method, such as centrifugation. The concentrated bacteria can then be used directly as a crude preparation, or further processing steps, well known to those skilled in the art, can be carried out, such as freeze-drying, dehydration, filtration, purification, optional freezing followed by thawing, sterilization, column chromatography, grinding, etc.

[0046] Preferably, the bacterial extract is a bacterial lysate comprising, in particular, the compounds contained within the bacterium, and therefore present in the bacterial pellet of the bacterial culture after centrifugation. Preferably, the bacterial extract is a bacterial lysate comprising the intracellular medium and / or the constituents of the cell walls and / or membranes.

[0047] A lysate commonly refers to a substance obtained at the end of the destruction or dissolution of biological cells by a phenomenon called cell lysis, which therefore causes the release of intracellular biological constituents naturally contained in the cells of the microorganism concerned.

[0048] For the purposes of the present invention, the term lysate is used interchangeably to designate the entire lysate obtained by the lysis of the microorganism concerned or only a fraction thereof.

[0049] The lysate used is therefore formed in whole or in part from intracellular biological constituents and from constituents of cell walls and membranes.

[0050] This cell lysis can be achieved through various technologies, such as osmotic shock, thermal shock such as freezing optionally followed by thawing, ultrasound, or under mechanical stress such as centrifugation, for example.

[0051] Preferably, the lysate is obtained by a process comprising the following steps: i. a centrifugation step; ii. a step of recovering the pellet; iii. a step of freezing the pellet; iv. a step of thawing the pellet; v. optionally, a sterilization step by autoclaving, in particular at 121 °C.

[0052] In a preferred embodiment of the invention, the bacterial extract comprises an inactivated bacterium.

[0053] Here, "inactivated" has the meaning commonly used by those skilled in the art, namely, the suppression of bacterial activity by various means (thermal, chemical, mechanical, enzymatic). The bacteria are preferably inactivated by heat, more preferably by autoclaving.

[0054] Preferably, the bacterium of the extract is Sphingomonas xenophaga. In a more preferred embodiment, the bacterium is a Sphingomonas xenophaga registered in accordance with the Treaty of Budapest, on 21 November 2019, with the National Collection of Microorganism Culture ((CNCM), Paris, France) under the number CNCM 1-5455 by L'Oréal, 101 Avenue Gustave Eiffel, 37390 Notre Dame d'Oé.

[0055] As examples of commercial Sphingomonas ferment extract products useful in the composition according to the present invention, one can cite that sold under the name of MEXORYL® SDT by the company NOVEAL.

[0056] Advantageously, the Sphingomonas ferment extract is present in the composition of the present invention in an amount ranging from 0.1 ppm to 100,000 ppm, preferably from 1 ppm to 10,000 ppm, more preferably from 5 ppm to 5,000 ppm, and even more preferentially from 10 ppm to 2500 ppm, relative to the total weight of the composition.

[0057] Advantageously, the weight ratio between the microorganism of species of the genus Bifidobacterium and the ferment extract of Sphingomonas is at least 10:1, preferably from 15:1 to 200:1, more preferably from 20:1 to 150:1. Compound of formula (I)

[0058] Preferably, the composition of the present invention comprises at least one compound of formula (I): year:

[0059] in which: Ri=H or -CH3 ; R2=H or -CH3 ; R3=-CH3 ; and Ri and R2 are not simultaneously equal to H.

[0060] The preferred compounds of formula (I) are madecassoside (compound of formula (I) with Ri=R3 =-CH3 and R2=H) and terminoloside (compound of formula (I) with Ri=H and R2=R3=-CH3).

[0061] These compounds of formula (I), and in particular their mixtures, can in particular be extracted from Centella asiatica according to the process described in application WO 2004 / 062678.

[0062] According to a preferred embodiment of the invention, a mixture of madecassoside and terminoloside is used. More preferably, the mixture of madecassoside and terminoloside has a madecassoside content ranging from 30% by weight to 70% by weight, in particular ranging from 45% by weight to 55% by weight, and more particularly 50% by weight, relative to the total weight of the mixture.

[0063] According to a particularly preferred embodiment of the invention, an extract of Centella asiatica comprising more than 95% by weight of the madecassoside / terminoloside mixture, relative to the weight of the extract, will be used.

[0064] An extract of Centella asiatica comprising more than 95% by weight of the madecassoside / terminoloside mixture (50 / 50 by weight) is sold in particular under the trade name Madecassoside by Bayer.

[0065] A commercial madecassoside product is also available from the company SEPPIC, which consists of madecassoside.

[0066] Advantageously, the compound of formula (I) described above is present in the composition in an amount ranging from 0.1 ppm to 100,000 ppm, preferably from 1 ppm to 10,000 ppm, more preferably from 5 ppm to 5,000 ppm, and even more preferably from 10 ppm to 2,500 ppm, relative to the total weight of the composition. Bisabolol

[0067] Preferably, the composition of the present invention comprises bisabolol.

[0068] Advantageously, bisabolol is present in the composition of the present invention in an amount ranging from 0.1 ppm to 50,000 ppm, preferably from 0.1 ppm to 10,000 ppm, more preferably from 0.15 ppm to 5,000 ppm, and even more preferably from 0.2 ppm to 1,000 ppm, relative to the total weight of the composition. Panthenol

[0069] Preferably, the composition of the present invention comprises panthenol.

[0070] Advantageously, panthenol is present in the composition of the present invention in an amount ranging from 0.001% by weight to 30% by weight, preferably from 0.01% by weight to 20% by weight, more preferably from 0.03% by weight to 15% by weight, even more preferably from 0.1% by weight to 10% by weight, relative to the total weight of the composition. Aqueous phase

[0071] The composition of the present invention may include an aqueous phase.

[0072] Preferably, the aqueous phase comprises water.

[0073] Advantageously, water is present in the composition of the present invention in an amount ranging from 50% by weight to 99.9% by weight, preferably from 70% by weight to 99.7% by weight, relative to the total weight of the composition.

[0074] Optionally, the aqueous phase comprises an organic solvent miscible with water (at room temperature at 25 °C) selected from monoalcohols, glycols and polyols having from 2 to 20 carbon atoms, such as octyldodecanol, glycerin, propylene glycol, butylene glycol, pentylene glycol, hexylene glycol, caprylyl glycol, dipropylene glycol, diethylene glycol; and mixtures thereof.

[0075] Advantageously, the aqueous phase is present in the composition of the present invention in an amount ranging from 60% by weight to 99.9% by weight, preferably from 70% by weight to 99.7% by weight, relative to the total weight of the composition. Additional cosmetic active ingredients

[0076] The composition of the present invention may include an additional cosmetic active ingredient in addition to the cosmetic active ingredients as defined above.

[0077] A person skilled in the art can adjust the type and quantity of additional cosmetic active ingredients according to the end use of the composition according to the present invention. Additional adjuvants or additives

[0078] The composition of the present invention may also contain conventional cosmetic adjuvants or additives, for example perfumes, chelating agents, preservatives and bactericides, surfactants, thickeners, pH correctors and mixtures thereof.

[0079] A person skilled in the art can choose the quantity of additional adjuvants or additives so that it does not have a negative impact on the final use of the composition according to the present invention.

[0080] According to a particularly preferred embodiment, the present invention proposes a composition comprising, relative to the total weight of the composition: a. from 0.01% by weight to 10% by weight of a Bifida ferment lysate; b. from 1 ppm to 2500 ppm of an extract of Salvia miltiorrhiza; And c. from 10 ppm to 2500 ppm of a Sphingomonas ferment extract. pharmaceutical form and process

[0081] The composition of the present invention may be in the form of an emulsion (O / W or W / O), or other liquid formats, for example toners, serums, lotions, creams, masks, sprays.

[0082] The composition of the present invention can be used for the care of keratinous materials, such as skin. In particular, the composition of the present invention can provide benefits to the skin, for example, repairing the skin and / or providing a soothing effect to the skin.

[0083] According to the second aspect, the present invention proposes a non-therapeutic method for the treatment of keratinous materials, comprising the application of the composition according to the first aspect of the present invention to the keratinous materials.

[0084] In some embodiments, the keratinous material is the skin. EXAMPLES

[0085] The following examples are given by way of non-limiting illustrations of the present invention.

[0086] The main raw materials used, their trade names and suppliers are listed in Table 1.

[0087] [Tables 1] INCI Name Trade Name Supplier Bifida Ferment Lysate Repair Complex CLR® PF CLR (Bifida ferment lysate) Sphingomonas ferment extract MEXORYL® SDT NOVEAL Salvia miltiorrhiza root extract NEUROFENSE MC SILAB Madecassoside MADECASSOSIDE SERDEX (SEPP IC)

[0088] Examples 1 to 3 of the invention and Comparative Examples 1 and 2

[0089] The compositions of the examples of the invention (El) 1 to 3 and of the comparative examples (EC) 1 and 2 were prepared on the basis of the quantities of components given in Table 2. The quantities are given by weight in ppm or percentage of each component relative to the total weight of the composition, MA meaning active material.

[0090] [Tables2] Components ELI EI.2 EL3 EC.l EC.2 Bifi da ferment lysate (%) 9.86 (MA) 9.86 (MA) 9.86 (MA) 9.86 (MA) 0 Sp hingomonas ferment extract (ppm) 880 (MA) 880 (MA) 880 (MA) 0 880 (MA) Salvi a root extract miltiorrhiza (ppm) 75 (MA) 75 (MA) 75 (MA) 0 75 (MA) Bisabolol (ppm) 0 10 10 0 0 Madecassoside (ppm) 0 1100 1100 0 0 Panthenol (%) 0 0 5 (MA) 0 0 Water QS100 QS100 QS100 QS100 QS100

[0091] The compositions of examples 1 to 3 of the invention represent the composition according to the present invention.

[0092] The composition of comparative example 1 does not include at least one extract from S al via miltiorrhiza and a Sphingomonas ferment extract.

[0093] The composition of comparative example 2 does not include at least one extract of Bifida ferment.

[0094] Preparation process:

[0095] The compositions of Examples 1 to 3 of the invention and of comparative examples 1 and 2 were prepared as follows: slowly add to water, where appropriate, Bifida ferment lysate, Sphingomonas ferment extract, Salvia miltiorrhiza root extract, bisabolol, madecassoside and panthenol in the corresponding amounts of Table 2, under stirring and at room temperature, to obtain a homogeneous mixture. Assessment

[0096] The soothing and repairing effects of the compositions of Examples 1 and 2 of the invention and of comparative Examples 1 and 2 were evaluated by detecting the levels of the inflammatory mediator (PGE2) and involucrin (IVL) on a 3D epidermal skin model stimulated with sodium dodecyl sulfate (SLS). More specifically, the soothing efficacy of the samples was evaluated by detecting the change in the inflammatory mediator (PGE2) level by ELISA, and the repairing efficacy of the samples was evaluated by detecting the change in the involucrin (IVL) level by immunofluorescence. Furthermore, the composition of Example 3 of the invention was evaluated in terms of soothing efficacy by also detecting the change in the inflammatory mediator (PGE2) level by ELISA.

[0097] In particular, the test was carried out as follows.

[0098] 1) Preparation of the solution:

[0099] The working solutions were prepared according to the test groups listed in Table 3.

[0100] [Tables3] Groups Sample Name Concentration Stimulation Condition Blank Control (BC) / / / Negative Control (NC) / / 0.1% SLS Positive Control 1 (PC 1) Dexamethasone 0.01% Positive Control 2 (PC 2) WY14643 50 µM Sample EC.1 Concentrations of Bifida ferment lysate, Sphinx ferment extract, Salvia miltiorrhiza root extract, Bisabolol, Madecass EC.2 ELI EI.2 EI.3 oside and panthenol are identical to those indicated in Table 2

[0101] 2) Administration i. Depending on the test groups, the models were transferred to a 6-well plate (previous addition of 0.9 ml of growth medium). ii. 25 µl of 0.1% SLS solution were added to the skin surface in the NC group, PCI group, PC2 group and sample group, and then incubated for 30 min. iii. After incubation, the corresponding concentration of the working solution was added to the PCI group and the PC2 group under liquid, the sample working solution was added uniformly to the skin surface, and then all models were incubated in a CO2 incubator (37 °C, 5% CO2) for 24 h. iv. After incubation, the test substance was washed with sterile PBS and the inserts were dried with a sterile cotton swab.

[0102] 3) Detection by ELISA

[0103] After incubation, the culture medium was collected in centrifuge tubes. The samples for ELISA detection were placed in a refrigerator at -80 °C until testing. Detection and analysis were performed according to the instructions in the operating procedure for each ELISA kit.

[0104] 4) Detection by immunofluorescence

[0105] The models used for immunofluorescence detection were sampled and fixed with 4 wt% paraformaldehyde. After fixation for 24 h, immunofluorescence detection was performed. The images were taken for microscopic observation and analysis.

[0106] The improvement rate and the inhibition rate were calculated according to the following equations:

[0107] Improvement rate (%) = Sample group - Negative control group x 1 QQ % • Negative control group

[0108] Inhibition rate (%) = Negative control group - Sample group x 1 QQ % Negative control group

[0109] GraphPad Prism software was used to analyze the data. Comparisons between groups were performed using the t-test. The statistical analysis was two-sided. P < 0.05 was considered a significant difference, and P < 0.01 was considered highly significant.

[0110] The results of the detection of PGE2 content have been summarized in Table 4. [YES] [Tables4] Groups P-value Inhibition rate (vs NC) BC / / NC 0.000 ## / PCI 0.000 ** 83.95% EC.1 0.021 * 15.77% EC.2 0.037 * 10.38% ELI 0.002 ** 27.04% EI.2 0.001 ** 28.96% EI.3 0.003 ** 30.90%

[0112] Note: Compared to group BC, value P<0.05#, value P<0.01##. Compared to group NC, value P<0.05*, value P<0.01**.

[0113] Table 4 shows that, compared to samples EC.1 and EC.2, samples ELI at 3 have a significantly improved calming effect.

[0114] The results of the immunofluorescence analysis of IVL have been summarized in Table 5.

[0115] [Tableaux5] Groups Value P Improvement Rate (vs NC) BC / / NC 0.003 ## / PC2 0.002 ** 47.62% EC.1 0.007 ** 36.51% EC.2 0.008 ** 49.21% ELI 0.000 ** 96.83% EI.2 0.001 ** 144.44%

[0116] Note: Compared to group BC, value P<0.05#, value P<0.01##. Compared to group NC, value P<0.05*, value P<0.01**.

[0117] Table 5 shows that, compared to samples EC.1 and EC.2, samples ELI and EI.2 have a significantly improved repair effect.

[0118] [Example 4 of the invention and Comparative Examples 3 and 41

[0119] Compositions of the example of the invention (E1) 4 and of the comparative examples (E1) 3 and 4 were prepared on the basis of the quantities of components given in Table 6. Quantities are given by weight in ppm or percentage of each component relative to the total weight of the composition, MA meaning active substance.

[0120] [Tableauxô] Components EL4 EC.3 EC.4 Bifida ferment lysate (%) 0.31 (MA) 0.31 (MA) 0 Sphingomonas ferment extract (ppm) 27.50 (MA) 0 27.50 (MA) Salvia miltiorrhiza root extract (ppm) 2.34 (MA) 0 2.34 (MA) Water (%) QS100 QS100 QS100

[0121] The composition of example 4 of the invention represents the composition according to the present invention.

[0122] The composition of comparative example 3 does not include at least one extract of S al via miltiorrhiza and one extract of ferment from Sphingomonas.

[0123] The composition of comparative example 4 does not include at least one extract of Bifida ferment.

[0124] Preparation process:

[0125] The compositions of Example 4 of the invention and comparative examples 3 and 4 were prepared as follows: slowly add to water the Bifida ferment lysate, the Sphingomonas ferment extract and the Salvia miltiorrhiza root extract (if applicable) according to the corresponding amounts in Table 6, under stirring and at room temperature, to obtain a homogeneous mixture. Assessment

[0126] The restorative effect of the compositions of example 4 of the invention and comparative examples 3 and 4 above was evaluated as follows. Repairing effect

[0127] The restorative effect of the samples of example 4 of the invention and comparative examples 3 and 4 was evaluated by detecting the change in Claudin-1 content on the basis of keratinocytes.

[0128] The cells used in this assay were keratinocytes.

[0129] In particular, the test was carried out as follows.

[0130] 1) Inoculation: After thawing the cells, when the growth rate Cellular growth reached approximately 60%, the cells were seeded in a 24-well plate and incubated in a CO2 incubator (37°C, 5% CO2) for 24 h.

[0131] 2) Preparation of the solution:

[0132] The working solutions were prepared according to the test groups listed in Table 7.

[0133] [Tables?] Groups Sample Name Concentration Blank Control (BC) / / Positive Control (PC) WY14643 50 qM Sample EC.3 The concentrations of Bifida ferment lysate, Sphingomon as ferment extract, and Salvia miltiorrhiza root extract are identical to those indicated in Table 6 EC.4 EL 4

[0134] 3) Administration: when the cell growth rate in the plaque is 24 When 40–60% of the wells were filled, three replications were administered to each group. The blank control group was treated with 1 ml of culture medium in each well, the positive control group with 1 ml of culture medium containing WY14643 in each well, and the sample group with 1 ml of culture medium containing the corresponding samples in each well. After administration, the 24-well plate was incubated in an incubator (37 °C, 5% CO2) for 24 hours.

[0135] 4) Detection by immunofluorescence: After incubation, the cells were fixed with 4% paraformaldehyde by weight for 30 min. After fixation, detection by immunofluorescence was performed. Images were taken for microscopic observation and analysis.

[0136] The rate of improvement was calculated according to the following equation:

[0137] Improvement Rate (%) = Sample - Control Group (blank) x ] QQ % control group

[0138] GraphPad Prism software was used to analyze the data. Comparisons between groups were performed using the t-test. The statistical analysis was two-sided. P < 0.05* was considered a significant difference, and P < 0.01** was considered highly significant.

[0139] The results of the Claudine-1 immunofluorescence analysis have been summarized in Table 8.

[0140] [Tables8] Groups Value P Improvement Rate (vs BC) BC / / PC 0.003 ** 67.00% EC.3 0.018 * 29.00% EC.4 0.004 ** 60.00% EI.4 0.001 ** 117.00%

[0141] It can be seen from Table 8 that, compared to samples EC.3 and EC.4, sample EI.4 significantly improved the repairing effect.

Claims

Demands

1. Composition, preferably for the care of keratinous materials, comprising: a) at least one microorganism of the species of the genus Bifidobacterium, the microorganism being in lysate form, b) at least one extract of Salvia miltiorrhiza, and c) an extract of ferment of Sphingomonas.

2. Composition according to claim 1, wherein the microorganism of the species of the genus Bifidobacterium is selected from the following species: Bifidobacterium longum, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium animalis, Bifidobacterium lactis, Bifidobacterium infantis, Bifidobacterium adolescentis, Bifidobacterium pseudobacterium, and mixtures thereof; preferably the microorganism is Bifidobacterium longum.

3. Composition according to claim 1 or 2, wherein the microorganism of species of the genus Bifidobacterium is present in an amount from 0.001% by weight to 20% by weight, preferably from 0.01% by weight to 15% by weight, more preferably from 0.01% by weight to 10% by weight, relative to the total weight of the composition.

4. Composition according to any one of claims 1 to 3, wherein the extract of Salvia miltiorrhiza is present in an amount from 0.1 ppm to 50,000 ppm, preferably from 0.1 ppm to 10,000 ppm, more preferably from 0.5 ppm to 5,000 ppm, and even more preferably from 1 ppm to 2,500 ppm, relative to the total weight of the composition.

5. Composition according to any one of claims 1 to 4, wherein the Sphingomonas ferment extract is present in an amount from 0.1 ppm to 100,000 ppm, preferably from 1 ppm to 10,000 ppm, more preferably from 5 ppm to 5,000 ppm, and even more preferably from 10 ppm to 2,500 ppm, relative to the total weight of the composition.

6. Composition according to claim 1, comprising, in relation to the total weight of the composition: a) from 0.01% by weight to 10% by weight of a Bifida ferment lysate;

7. b) from 1 ppm to 2500 ppm of an extract of Salvia miltiorrhiza; And (c) 10 ppm to 2500 ppm of a Sphingomonas ferment extract. Composition according to any one of claims 1 to 6, further comprising a compound of formula (I):

8.

9.

10. in which: Ri=H or -CH3; R2=H or -CH3; R3=-CH3; and Riet R2 are not simultaneously equal to H. Composition according to any one of claims 1 to 7, further comprising bisabolol, preferably in an amount from 0.1 ppm to 50,000 ppm, preferably from 0.1 ppm to 10,000 ppm, more preferably from 0.15 ppm to 5,000 ppm, even more preferably from 0.2 ppm to 1,000 ppm, relative to the total weight of the composition. Composition according to any one of claims 1 to 8, further comprising panthenol, preferably in an amount from 0.001% by weight to 30% by weight, more preferably from 0.01% by weight to 20% by weight, even more preferably from 0.03% by weight to 15% by weight, most preferably from 0.1% by weight to 10% by weight, relative to the total weight of the composition. Non-therapeutic method for the treatment of keratinous materials, comprising the application of the composition according to any one of claims 1 to 9 on keratinous materials.