Composition and method for caring for keratin materials
A composition with Bifidobacterium lysate, Salvia miltiorrhiza, and Sphingomonas ferment extract addresses skin aging issues by enhancing repair and soothing effects, improving skin health through reduced inflammation and increased repair markers.
Patent Information
- Authority / Receiving Office
- WO · WO
- Patent Type
- Applications
- Current Assignee / Owner
- Filing Date
- 2024-09-27
- Publication Date
- 2026-04-02
AI Technical Summary
Existing cosmetic compositions fail to deliver satisfactory repairing and soothing effects on the skin, particularly due to decreases in collagen content, keratinocyte differentiation, and synthesis of glycosaminoglycans, leading to skin aging and damage.
A composition comprising Bifidobacterium species lysate, Salvia miltiorrhiza extract, and Sphingomonas ferment extract is formulated to provide repairing and soothing benefits to keratin materials.
The composition effectively repairs and soothes keratin materials by reducing inflammatory mediators and enhancing skin repair markers, demonstrating significant improvement in skin health.
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Figure PCTCN2024121604-FTAPPB-I100001 
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Figure PCTCN2024121604-FTAPPB-I100003
Abstract
Description
COMPOSITION AND METHOD FOR CARING FOR KERATIN MATERIALSTECHNICAL FIELD
[0001] The present invention relates to a cosmetic composition. In particular, the present invention relates to a composition for caring for keratin materials. The present invention also relates to a non-therapeutic method for caring for keratin materials.BACKGROUND ART
[0002] The skin is the protective barrier for the human body. It protects the interior of the body from physical injury (such as trauma) and biological injury (such as bacteria, viruses or fungi) . The epidermis is a keratinized stratified pavimentous epithelium. Its mean thickness ranges from 60 to 100μm and may reach 600 to 700μm on the sole of the feet and the palm of the hands. It consists mainly of keratinocytes, but also other cells, and rests on a basal membrane that separates it from the dermis.
[0003] The main changes concern the dermis and are a decrease in the collagen content and in the thickness of the dermis.
[0004] The main changes concerning the epidermis are a decrease in keratinocyte differentiation, resulting in a deficit in the proteins matrix of the cornified cell, an increase in metalloproteinases, which are proteases that degrade the extracellular matrix and that participate in ageing of the skin, and also a decrease in the synthesis of various glycosaminoglycans.
[0005] The development of formulations dedicated to caring for the skin is permanent.
[0006] A wide variety of cosmetic compositions have been used to care for the skin, for example, to deliver a repairing and soothing effect to the skin. However, for most of them, the effect on caring for the skin, especially repairing and soothing the skin is not satisfying.
[0007] Thus, there is still a need to formulate a composition for caring for the skin, which can deliver good repairing and / or soothing effect.SUMMARY OF THE INVENTION
[0008] An object of the present invention is thus to develop a composition for caring for keratin materials, which can deliver good repairing and / or soothing effect.
[0009] Another object of the present invention is to provide a cosmetic process for caring for keratin materials.
[0010] Accordingly, in a first aspect, the present invention provides a composition, preferably for caring for keratin materials, comprising:
[0011] a) at least one microorganism of the genus Bifidobacterium species, the microorganism being in the lysate form,
[0012] b) at least one Salvia miltiorrhiza extract, and
[0013] c) Sphingomonas ferment extract.
[0014] In a second aspect, the present invention provides a non-therapeutic method for caring for keratin materials, comprising applying the composition according to the first aspect of the present invention to the keratin materials.
[0015] The composition of the present invention can deliver good repairing and / or soothing effect to the keratin materials.
[0016] Other subjects and characteristics, aspects and advantages of the present invention will be set forth in the description that follows, and in part, will be obvious from the description, or may be learned by practice of the present invention.DETAILED DESCRIPTION OF THE INVENTION
[0017] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art the present invention belongs to. When the definition of a term in the present description conflicts with the meaning as commonly understood by those skilled in the art the present invention belongs to, the definition described herein shall apply.
[0018] In that which follows and unless otherwise indicated, the limits of a range of values are included within this range, in particular in the expressions "between... and…" and "from... to... " .
[0019] Moreover, the expression "at least one" used in the present description is equivalent to the expression "one or more" .
[0020] Throughout the instant application, the term “comprising” is to be interpreted as encompassing all specifically mentioned features as well as optional, additional, unspecified ones. As used herein, the use of the term “comprising” also discloses the embodiment wherein no features other than the specifically mentioned features are present (i.e. “consisting of” ) .
[0021] Unless otherwise specified, all numerical values expressing amount of ingredients and the like which are used in the description and claims are to be understood as being modified by the term “about” . Accordingly, unless indicated to the contrary, the numerical values and parameters described herein are approximate values which are capable of being changed according to the desired purpose as required.
[0022] For the purposes of the present invention, the term "keratin materials" is intended to cover human skin, including facial skin and the lips. Facial skin is most particularly considered according to the present invention.
[0023] All percentages in the present invention refer to weight percentage, unless otherwise specified.
[0024] According to the first aspect, the composition of the present invention comprises:
[0025] a) at least one microorganism of the genus Bifidobacterium species, the microorganism being in the lysate form,
[0026] b) at least one Salvia miltiorrhiza extract, and
[0027] c) Sphingomonas ferment extract.
[0028] Microorganism of the genus Bifidobacterium species
[0029] According to the first aspect, the composition of the present invention comprises at least one microorganism of the genus Bifidobacterium species, the microorganism being in the lysate form.
[0030] A lysate commonly denotes a material obtained after the destruction or the dissolution of biological cells by a phenomenon referred to as cell lysis, thus causing the release of the intracellular biological constituents naturally contained in the cells of the microorganism in question.
[0031] In the context of the present invention, the term "lysate" is used without preference to denote all the lysate obtained by lysis of the microorganism in question or only a fraction of the lysate. The lysate used is thus formed completely or partially of the intracellular biological constituents and of the constituents of the walls and cell membranes.
[0032] More specifically, the lysate contains the cellular cytoplasmic fraction containing enzymes, such as lactic acid dehydrogenase, phosphatases, phosphoketolases or transaldolases, and the metabolites. By way of illustration, the constituents of the cell walls are in particular peptidoglycan, murein or mucopeptide and teichoic acid, and the constituents of the cell membranes are composed of glycerophospholipids.
[0033] This cell lysis can be carried out by various techniques, for example by osmotic shock, by thermal shock, by ultrasound, or under a mechanical stress, for example by centrifugation. More particularly, this lysate may be obtained according to the technology described in patent US 4 464 362 and especially according to the following protocol.
[0034] The microorganism (s) of the genus Bifidobacterium species are preferably cultured under anaerobiosis in an appropriate culture medium, for example according to the conditions described in the documents US 4 464 362 and EP 4376804. When 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℃, for 30 minutes. The microorganism (s) thus formed are subsequently collected by a conventional separation technique, such as membrane filtration or centrifugation, and are then re-suspended in a sterile physiological solution of NaCl.
[0035] The lysate of these microorganisms can subsequently be obtained by ultrasonic disintegration of the solution obtained above, so as to release the cytoplasmic fractions, the cell wall fragments and the products resulting from metabolism. All the components, in their natural distribution, are subsequently stabilized in a weakly acidic aqueous solution. The total content of active material (s) , present in the lysate obtained by this process, preferably ranges from 0.1%to 50%by weight, more preferentially from 1%to 20%by weight, and better still this content is 5%by weight, relative to the total weight of the lysate.
[0036] The lysate can be used in various forms, in particular in the form of a solution or in pulverulent form.
[0037] Preferably, the microorganism of the genus Bifidobacterium species 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.
[0038] More preferably, the microorganism of the genus Bifidobacterium species is Bifidobacterium longum.
[0039] Mention may in particular be made, as lysate capable of being used within the meaning of the invention, of the ingredient having the INCI name: Bifida Ferment Lysate.
[0040] Mention may in particular be made, by way of example, of the product sold under the name Repair Complex by K. Richter GmbH, and which is formed of an inactivated lysate of the species Bifidobacterium longum.
[0041] Advantageously, the microorganism of the genus Bifidobacterium species is present in the composition of the present invention in an amount ranging from 0.001 wt. %to 20 wt. %, preferably from 0.01 wt. %to 15 wt. %, more preferably from 0.01 wt. %to 10 wt. %, relative to the total weight of the composition.
[0042] Salvia miltiorrhiza extracts
[0043] According to the first aspect, the composition according to the present invention comprises at least one Salvia miltiorrhiza extract.
[0044] Preferably, the Salvia miltiorrhiza extract is selected from salvia miltiorrhiza root extract and Salvia miltiorrhiza leaf extract.
[0045] More preferably, the composition of the present invention comprises a Salvia miltiorrhiza root extract.
[0046] Advantageously, the Salvia miltiorrhiza root extract comprises at least 5 wt. %of total polyphenols relative to the total weight of dry matter of the extract, preferably from 5 wt. %to 15 wt. %of total polyphenols, relative to the total weight of dry matter of the extract.
[0047] Mention may in particular be made, by way of example, of the product sold under the name NEUROFENSE MC by SILAB, which is Salvia miltiorrhiza root extract.
[0048] Advantageously, the Salvia miltiorrhiza extract is present in the composition of the present invention in an amount ranging from 0.1 ppm to 50000 ppm, preferably from 0.1 ppm to 10000 ppm, more preferably from 0.5 ppm to 5000 ppm, and even more preferably from 1 ppm to 2500 ppm, relative to the total weight of the composition.
[0049] Sphingomonas ferment extract
[0050] According to the first aspect, the composition of the present invention comprises Sphingomonas ferment extract.
[0051] 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 popular for its reduction of lipid peroxidation and functioning of the key biomarkers of senescence, thus preventing the skin aging.
[0052] The extracts according to the invention can be prepared by any conventional method well known to persons skilled in the art. These methods typically comprise a step of fermentation, isolation, separation, and purification.
[0053] Typically, the bacterium of Sphingomonas genus is cultivated in a suitable medium and cultivated at 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 raw preparation, or additional steps of treatment well known to person skilled in the art can be carried out, such as lyophilization, dehydration, filtration, purification, freezing optionally followed by a thawing, sterilization, column chromatography, grinding, etc.
[0054] Preferably, the bacterial extract is a lysate of bacteria comprising in particular the compounds comprised in the bacterium, therefore present in the bacterial pellet of the bacterial culture after centrifugation. Preferably, the bacterial extract is a lysate of bacteria comprising the intracellular medium and / or the constituents of the walls and / or cell membranes.
[0055] A lysate commonly designates a material obtained at the end of the destruction or dissolution of biological cells by a phenomenon called cell lysis thus causing the release of the intracellular biological constituents naturally contained in the cells of the microorganism considered.
[0056] In terms of the present invention, the term lysate is used indifferently to designate all the lysate obtained by lysis of the microorganism concerned or only a fraction of the latter.
[0057] The lysate implemented is therefore form entirely or partially of the intracellular biological constituents and constituents of the cell walls and membranes.
[0058] This cellular lysis can be carried out using different technologies, such as for example osmotic shock, thermal shock such as a freezing optionally followed by a thawing, by ultrasound, or under mechanical stress of the centrifugation type for example.
[0059] Preferably, the lysate is obtained by a method comprising the following steps:
[0060] i) a step of centrifugation;
[0061] ii) a step of recovering the pellet;
[0062] iii) a step of freezing the pellet;
[0063] iv) a step of thawing the pellet;
[0064] v) optionally a step of sterilizing via autoclaving, in particular at 121℃.
[0065] In a preferred embodiment of the invention the extract of bacteria comprises an inactivated bacterium.
[0066] Here, "inactivated" has the meaning that is commonly used by persons skilled in the art, i.e. the suppression of the activity of the bacterium under the effect of various causes (heat, chemical, mechanical, enzyme substances) . Preferably, the bacterium is inactivated by heat, more preferably by autoclaving.
[0067] Preferably, the bacterium of the extract is Sphingomonas xenophaga. In a more preferred embodiment, the bacterium is a Sphingomonas xenophaga registered according to the Budapest Treaty, on November 21, 2019, with the Collection Nationale de Culture de Microorganismes ( (CNCM) , Paris, France) under the number CNCM I-5455by L’Oréal, 101 Avenue Gustave Eiffel, 37390 Notre Dame d’Oé.
[0068] As examples for commercial products of sphingomonas ferment extract useful in the composition according to the present invention, mention can be made to that sold under the name SDT by the company NOVEAL.
[0069] Advantageously, the Sphingomonas ferment extract is present in the composition of the present invention in an amount ranging from 0.1 ppm to 100000 ppm, preferably from 1 ppm to 10000 ppm, more preferably from 5 ppm to 5000 ppm, and even more preferably from 10 ppm to 2500 ppm, relative to the total weight of the composition, relative to the total weight of the composition.
[0070] Advantageously, the weight ratio of the microorganism of the genus Bifidobacterium species to the Sphingomonas ferment extract is at least 10: 1, preferably from 15: 1 to 200: 1, more preferably from 20: 1 to 150: 1.
[0071] Compound of formula (I)
[0072] Preferably, the composition of the present invention comprises at least one compound of formula (I) :
[0073] wherein: R1=H or -CH3; R2=H or -CH3; R3=-CH3; and R1 and R2 do not simultaneously equal to H.
[0074] The preferred compounds of formula (I) are madecassoside (compound of formula (I) with R1=R3=-CH3 and R2=H) and terminoloside (compound of formula (I) with R1=H and R2=R3=-CH3) .
[0075] These compounds of formula (I) and in particular their mixtures can especially be extracted from Centella asiatica according to the process described in Application WO 2004 / 062678.
[0076] According to a preferred embodiment of the invention, use is made of a mixture of madecassoside and of terminoloside. More preferably, the mixture of madecassoside and of terminoloside has a madecassoside content ranging from 30 wt. %to 70 wt. %, particularly ranging from 45 wt. %to 55 wt. %, and more particularly of 50 wt. %, relative to the total weight of the mixture.
[0077] According to a particularly preferred embodiment of the invention, use will be made of a Centella asiatica extract comprising more than 95 wt. %of the madecassoside / terminoloside mixture, relative to the weight of the extract.
[0078] A Centella asiatica extract comprising more than 95 wt. %of madecassoside / terminoloside mixture (50 / 50 wt. %) is sold in particular under the trade name Madecassoside by Bayer.
[0079] A commercial product of madecassoside is also available from the company SEPPIC, which consists of madecassoside.
[0080] Advantageously, the compound of formula (I) described above is present in the composition in an amount ranging from 0.1 ppm to 100000 ppm, preferably from 1 ppm to 10000 ppm, more preferably from 5 ppm to 5000 ppm, and even more preferably from 10 ppm to 2500 ppm, relative to the total weight of the composition, relative to the total weight of the composition.
[0081] Bisabolol
[0082] Preferably, the composition of the present invention comprises bisabolol.
[0083] Advantageously, bisabolol is present in the composition of the present invention in an amount ranging from 0.1 ppm to 50000 ppm, preferably from 0.1 ppm to 10000 ppm, more preferably from 0.15 ppm to 5000 ppm, even more preferably from 0.2 ppm to 1000 ppm, relative to the total weight of the composition.
[0084] Panthenol
[0085] Preferably, the composition of the present invention comprises panthenol.
[0086] Advantageously, panthenol is present in the composition of the present invention in an amount ranging from 0.001 wt. %to 30 wt. %, preferably from 0.01 wt. %to 20 wt. %, more preferably from 0.03 wt. %to 15 wt. %, even more preferably from 0.1 wt. %to 10 wt. %, relative to the total weight of the composition.
[0087] Aqueous phase
[0088] The composition of the present invention may comprise an aqueous phase.
[0089] Preferably, the aqueous phase comprises water.
[0090] Advantageously, water is present in the composition of the present invention in an amount ranging from 50 wt. %to 99.9 wt. %, preferably from 70 wt. %to 99.7 wt. %, relative to the total weight of the composition.
[0091] Optionally, the aqueous phase comprises an organic solvent miscible with water (at room temperature 25℃) 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.
[0092] Advantageously, the aqueous phase is present in the composition of the present invention in an amount ranging from 60 wt. %to 99.9 wt. %, preferably from 70 wt. %to 99.7 wt. %, relative to the total weight of the composition.
[0093] Additional cosmetic active ingredients
[0094] The composition of the present invention may comprise an additional cosmetic active ingredient in addition to the cosmetic active ingredients as defined previously.
[0095] The skilled in the art can adjust the type and their amount of the additional cosmetic active ingredients based on the final use of the composition according to the present invention.
[0096] Additional adjuvants or additives
[0097] The composition of the present invention may also contain conventional cosmetic adjuvants or additives, for instance fragrances, chelating agents, preserving agents and bactericides, surfactants, thickeners, pH regulators, and mixtures thereof.
[0098] The skilled in the art can select the amount of the additional adjuvants or additive so as not to adversely impact the final use of the composition according to the present invention.
[0099] According to a particularly preferred embodiment, the present invention provides a composition comprising, relative to the total weight of the composition:
[0100] a) from 0.01 wt. %to 10 wt. %of Bifida ferment lysate;
[0101] b) from 1 ppm to 2500 ppm of Salvia miltiorrhiza extract; and
[0102] c) from 10 ppm to 2500 ppm of Sphingomonas ferment extract.
[0103] Galenic form and method
[0104] The composition of the present invention can be in the form of an emulsion (O / W or W / O) , or other liquid formats, for example, toners, serums, lotions, creams, masks, sprays.
[0105] The composition of the present invention can be used for caring for keratin materials, such as the skin. In particular, the composition of the present invention can bring benefits to the skin, for example repair the skin and / or deliver soothing effect to the skin.
[0106] According to the second aspect, the present invention provides a non-therapeutic method for caring for keratin materials, comprising applying the composition according to the first aspect of the present invention to the keratin materials.
[0107] In some embodiments, the keratin material is the skin.
[0108] EXAMPLES
[0109] The examples that follow are given as non-limiting illustrations of the present invention.
[0110] Main raw materials used, trade names and suppliers thereof are listed in Table 1.
[0111] Table 1
[0112] Invention Examples 1-3 and Comparative Examples 1-2
[0113] Compositions of invention examples (IE) 1-3 and comparative examples (CE) 1-2 were prepared based on the amounts of components given in Table 2. The amounts are given by weight in ppm or percentage of each component relative to the total weight of the composition, wherein AM stands for active material.
[0114] Table 2
[0115] Compositions of invention examples 1-3 represents the composition according to the present invention.
[0116] Composition of comparative example 1 does not comprise at least one Salvia miltiorrhiza extract and Sphingomonas ferment extract.
[0117] Composition of comparative example 2 does not comprise at least one Bifida ferment extract.
[0118] Preparation process:
[0119] Compositions of invention examples 1-3 and comparative example 1-2 were prepared as follows: adding, if any, Bifida ferment lysate, Sphingomonas ferment extract, Salvia miltiorrhiza root extract, bisabolol, madecassoside, and panthenol according to corresponding amounts in Table 2 slowly into water with stirring under room temperature to obtain a homogeneous mixture.
[0120] Evaluation
[0121] Soothing effect and repairing effect of compositions of invention examples 1-2 and comparative examples 1-2 were evaluated by detection of inflammatory mediator (PGE2) and Involucrin (IVL) contents based on 3D epidermal skin model stimulated by sodium dodecyl sulfate (SLS) . Specifically, the soothing efficacy of the samples was evaluated by detecting the change in the content of inflammatory mediator (PGE2) through ELISA, the repairing efficacy of the samples was evaluated by detecting the change in the contents of Involucrin (IVL) through Immunofluorescence. In addition, composition of invention examples 3 was evaluated in terms of the soothing efficacy by detecting the change in the content of inflammatory mediator (PGE2) through ELISA as well.
[0122] In particular, the test was carried out as follows.
[0123] 1) Solution preparation:
[0124] The working solutions were prepared according to the test groups listed in Table 3.
[0125] Table 3
[0126] 2) Administration
[0127] i) According to the test groups, the models were transferred to 6-well plate (added 0.9mL of Growth medium in advance) .
[0128] ii) 25μL of 0.1%SLS solution was added on the skin surface in NC group, PC1 group, PC2 group and sample group, then was incubated for 30 min.
[0129] iii) After incubation, the corresponding concentration of working solution was added to the PC1 group and PC2 group under liquid, the sample working solution was added evenly on the skin surface, and then all the models were incubated in CO2incubator (37℃, 5%CO2) for 24h.
[0130] iv) After incubation, washed the test substance with sterile PBS, and dried the inserts with sterile cotton swab.
[0131] 3) ELISA detection
[0132] After incubation, the culture medium was collected in centrifugal tubes. And the samples for ELISA detection were placed in a-80℃refrigerator until the test. The detection and analysis were carried out according to the operating instructions of each ELISA kit.
[0133] 4) Immunofluorescence detection
[0134] The models used for immunofluorescence detection were sampled and fixed with 4 wt. %paraformaldehyde. After fixation for 24h, immunofluorescence detection was performed. The images were taken for observation under the microscope and analyzed.
[0135] Improvement rate and inhibition rate was calculated according to the following equations:
[0136] GraphPad Prism software was used to analyze the data. The comparisons between groups were performed using t-test statistical analysis. Statistical analysis was two-tailed. P<0.05 was considered as significant difference, P<0.01 was considered as extremely significant difference.
[0137] The results of PGE2 content detection were summarized in Table 4.
[0138] Table 4
[0139] Note: Compared with BC group, P-value<0.05#, P-value<0.01##. Compared with NC group, P-value<0.05*, P-value<0.01**.
[0140] It can be seen from Table 4 that as compared with samples of CE. 1 and CE. 2, samples of IE. 1-3 have significantly improved soothing effect.
[0141] The results of IVL immunofluorescence analysis were summarized in Table 5.
[0142] Table 5
[0143] Note: Compared with BC group, P-value<0.05#, P-value<0.01##. Compared with NC group, P-value<0.05*, P-value<0.01**.
[0144] It can be seen from Table 5 that as compared with samples of CE. 1 and CE. 2, samples of IE. 1 and IE. 2 have significantly improved repairing effect.
[0145] Invention Example 4 and Comparative Examples 3-4
[0146] Compositions of invention example (IE) 4 and comparative examples (CE) 3-4 were prepared based on the amounts of components given in Table 6. The amounts are given by weight in ppm or percentage of each component relative to the total weight of the composition, wherein AM stands for active material.
[0147] Table 6
[0148] Compositions of invention example 4 represents the composition according to the present invention.
[0149] Composition of comparative example 3 does not comprise at least one Salvia miltiorrhiza extract and Sphingomonas ferment extract.
[0150] Composition of comparative example 4 does not comprise at least one bifida ferment extract.
[0151] Preparation process:
[0152] Compositions of invention example 4 and comparative example 3-4 were prepared as follows: adding Bifida ferment lysate, Sphingomonas ferment extract and Salvia miltiorrhiza root extract (if any) according to corresponding amounts in Table 6 slowly into water with stirring under room temperature to obtain a homogeneous mixture.
[0153] Evaluation
[0154] The repairing effect of compositions of invention example 4 and comparative examples 3-4 above was evaluated as follows.
[0155] Repairing effect
[0156] The repairing effect of the samples of invention example 4 and comparative examples 3-4 was evaluated by detecting the change in Claudin-1 content based on keratinocytes.
[0157] The cells used in this test was keratinocytes.
[0158] In particular, the test was carried out as follows.
[0159] 1) Inoculation: After the cells were thawed, when the cell laying rate reached about 60%, cells were seeded into 24-well plate, and incubated in CO2 incubator (37 ℃, 5%CO2) for 24h.
[0160] 2) Solution preparation:
[0161] The working solutions were prepared according to the test groups listed in Table 7.
[0162] Table 7
[0163] 3) Administration: when the cell laying rate in the 24-well plate reached40~60%, administration with 3 replications was performed in each group. Blank control group was added with 1mL culture medium in each well, positive control group with 1mL culture medium containing WY14643 in each well, and sample group with 1mL culture medium containing the corresponding samples in each well. After administration, the 24-well plate was incubated in incubator (37 ℃, 5%CO2) for 24h.
[0164] 4) Immunofluorescence detection: After incubation, the cells were fixed with 4 wt. %paraformaldehyde for 30 min. After fixation, performed immunofluorescence detection. The images were taken for observation under the microscope and analyzed.
[0165] Improvement rate was calculated according to the following equation:
[0166] GraphPad Prism software was used to analyze the data. The comparisons between groups were performed using t-test statistical analysis. Statistical analysis was two-tailed. P<0.05*was considered as significant difference, P<0.01**was considered as extremely significant difference.
[0167] The results of claudin-1 immunofluorescence analysis were summarized in Table 8.
[0168] Table 8
[0169] It can be seen from Table 8 that as compared with samples of CE. 3 and CE. 4, sample of IE.4 has significantly improved repairing effect.
Claims
1.A composition, preferably for caring for keratin materials, comprising:a) at least one microorganism of the genus Bifidobacterium species, the microorganism being in the lysate form,b) at least one Salvia miltiorrhiza extract, andc) Sphingomonas ferment extract.2.Composition according to claim 1, wherein the microorganism of the genus Bifidobacterium species 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 the genus Bifidobacterium species is present in an amount ranging from 0.001 wt. %to 20 wt. %, preferably from 0.01 wt. %to 15 wt. %, more preferably from 0.01 wt. %to 10 wt. %, relative to the total weight of the composition.4.Composition according to any of claims 1 to 3, wherein the Salvia miltiorrhiza extract is selected from Salvia miltiorrhiza root extract and Salvia miltiorrhiza leaf extract, preferably the composition comprises a Salvia miltiorrhiza root extract, preferably comprising at least 5 wt. %, preferably from 5 wt. %to 15 wt. %of total polyphenols, relative to the total weight of dry matter of the extract.5.Composition of any of claims 1 to 4, wherein the Salvia miltiorrhiza extract is present in an amount ranging from 0.1 ppm to 50000 ppm, preferably from 0.1 ppm to 10000 ppm, more preferably from 0.5 ppm to 5000 ppm, and even more preferably from 1 ppm to 2500 ppm, relative to the total weight of the composition.6.Composition of any of claims 1 to 5, wherein the sphingomonas ferment extract is present in an amount ranging from 0.1 ppm to 100000 ppm, preferably from 1 ppm to 10000 ppm, more preferably from 5 ppm to 5000 ppm, and even more preferably from 10 ppm to 2500 ppm, relative to the total weight of the composition.7.Composition of any of claims 1 to 6, wherein the weight ratio of the microorganism of the genus Bifidobacterium species to the Sphingomonas ferment extract is at least 10: 1, preferably from 15: 1 to 200: 1, more preferably from 20: 1 to 150: 1.8.Composition according to claim 1, comprising, relative to the total weight of the composition:a) from 0.01 wt. %to 10 wt. %of Bifida ferment lysate;b) from 1 ppm to 2500 ppm of Salvia miltiorrhiza extract; andc) from 10 ppm to 2500 ppm of Sphingomonas ferment extract.9.Composition according to any of claims 1 to 8, further comprising a compound of formula (I) : wherein: R1=H or -CH3; R2=H or -CH3; R3=-CH3; and R1 and R2 do not simultaneously equal H.10.Composition according to claim 9, wherein the compound of formula (I) is selected from madecassoside, terminoloside, and mixtures thereof.11.Composition according to claim 9 or 10, wherein the compound of formula (I) is present in an amount ranging from 0.1 ppm to 100000 ppm, preferably from 1 ppm to 10000 ppm, more preferably from 5 ppm to 5000 ppm, and even more preferably from 10 ppm to 2500 ppm, relative to the total weight of the composition.12.Composition according to any one of claims 1 to 11, further comprising bisabolol, preferably in an amount ranging from 0.1 ppm to 50000 ppm, preferably from 0.1 ppm to 10000 ppm, more preferably from 0.15 ppm to 5000 ppm, even more preferably from 0.2 ppm to 1000 ppm, relative to the total weight of the composition.13.Composition according to any one of claims 1 to 12, further comprising panthenol, preferably in an amount ranging from 0.001 wt. %to 30 wt. %, more preferably from 0.01 wt. %to 20 wt. %, even more preferably from 0.03 wt. %to 15 wt. %, most preferably from 0.1 wt. %to 10 wt. %, relative to the total weight of the composition.14.Composition according to any one of claims 1 to 13, further comprising water, preferably in an amount ranging from 50 wt. %to 99.9 wt. %, preferably from 70 wt. %to 99.7 wt. %, relative to the total weight of the composition.15.A non-therapeutic method for caring for keratin materials, comprising applying the composition according to any of claims 1 to 14 to the keratin materials.
Citation Information
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