FR3036962A1 - COSMETIC COMPOSITION BASED ON CENTELLA ASIATICA - Google Patents

Abstract

Cosmetic composition containing at least (a) Centella Asiatica cells or a crushed material of such cells, (b) an extract of Saussurea involucrata, and (c) a carrier for cosmetic application.

Description

The present invention relates to a cosmetic composition based on plant cells or cell extracts.

Many cosmetic compositions have already been proposed to act on the skin, in particular to ensure an energizing action on the skin. Even if these compositions had a certain activity, it was noticed that by using particular plant cells, it was possible to obtain an excellent cosmetic activity, in particular energizing for the skin. The subject of the invention is specifically a cosmetic composition containing at least (a) Centella Asiatica cells or a crushed material of such cells, (b) an extract of Saussurea involucrata, and (c) a support for cosmetic application. The composition according to the invention advantageously has one or more of the following characteristics, preferably a combination of such characteristics: the composition contains at least (a) dedifferentiated and eluted Centella Asiatica cells, preferably in the form of a ground meal, and (b) an extract of Saussurea involucrata. the composition contains at least (a) a crushed dedifferentiated and eluted Centella Asiatica cell, and (b) dedifferentiated and eluted Saussurea involucrata cells in the form of a crushed material of such cells. - The composition further contains an extract of cells selected from the family consisting of Araliaceae and Apiaceae, said extract being cosmetically acceptable. The composition also contains an extract of plant cells of the genus Panax from the family Araliaceae and / or the genus Angelica or Angelina from the Apiaceae family. the composition contains a plant extract chosen from Panax Pseudoginseng, Panax ginseng, Panax notoginseng, Angelica or Angelina Sinensis, and a mixture of such extracts. the composition contains an effective amount of Centella Asiatica cells or crushed such cells, and Saussurea involucrata extract, for its use to ensure an energizing cosmetic activity for the skin and / or the proteasomic activity of the skin. the composition contains from 0.02% to 10% by weight, preferably from 0.05 to 0.5% by weight, in particular from 0.06% to 0.2% by weight of Centella Asiatica cells, or crushing such cells, and from 0.02% to 10% by weight, preferably from 0.05 to 0.5% by weight, in particular from 0.06% to 0.2% by weight of Saussurea extract. 15 involucrata. the cells of Centella Asiatica are dedifferentiated and elicited cells grown in vitro. the composition contains a dedifferentiated and eluted Centella Asiatica ground material having a weight average size of less than 10 μm, in particular less than 5 μm, preferably less than - the composition is suitable for cleaning the skin. the composition is in the form of powder, creams, pastes, gels, suspensions, fluid or liquid compositions, solid or semi-solid compositions, sticks, sticks, pencils, solid or semi-solid gel with release The composition contains an effective amount of Centella Asiatica cells or crushed such cells, and Saussurea involucrata cell extract, for use in inducing stimulation of the basal cell respiration rate of non-permeabilized human keratinocytes. at least 5%, advantageously at least 20%, preferably at least 50%, in particular at least 75%, and / or 3036962 3 for its use induce stimulation of the mitochondrial respiration rate of permeabilized human keratinocytes at least 5%, advantageously at least 20%, preferably at least 50%, in particular at least 75%, in the presence of pyruvate-malate, and / or for its use to induce a stimulation of the synthesis rate of Adenisone triphosphate (ATP) of non-permeabilized human keratinocytes by at least 0.5%, advantageously by at least 10%, preferably by at least 20%, in particular by at least 50%, more particularly at least 75%, and / or for its use to induce a stimulation of the mitochondrial Adenisone Triphosphate (ATP) synthesis rate of permeabilized human keratinocytes by at least 0.5%, advantageously at least 10%, preferably at least 20%, in particular at least 50%, in the presence of pyruvate-malate, and / or for its use to induce stimulation of the sum (synthesis rate Adenisone triphosphate (ATP) cell + synthesis rate of cellular Adenosine monophosphate (AMP) + synthesis rate of Adenosine Cell Diphosphate (ADP)) of at least 10%, advantageously at least 20% preferably at least 50%, in particular at least 100% of non-permeate human keratinocytes after 5 days of contact, while still advantageously maintaining a ratio of the energy load between 0.74 and 0.85, and / or for its use to allow differentiation and proliferation of human keratinocytes in culture, and / or for its use to allow a better cohesion of the cells of the keratinocytes of the human skin, and / or for its use to ensure a vital effect of force for the skin, and / or for its use to ensure a moisturizing effect by NMF, and / or for its use to ensure a homeostasis effect, and / or for its use to ensure cellular cohesion of keratinocytes, and / or 3036962 4 for its use to ensure a cell renewal effect by proliferation and differentiation of cutaneous cells. The process for manufacturing dedifferentiated and eluted cells is advantageously a process of the type described in WO00 / 077881. In the present specification, dedifferentiated plant cells are any plant cell that does not exhibit any of the characteristics of a particular specialization and is capable of living on its own and not dependent on other cells. Dedifferentiated plant cells can be obtained from plant material derived from whole plant or part of plant such as leaves, stems, flowers, petals, roots, fruits, their skin, the envelope protecting them, 15 seeds, anthers, sap, thorns, buds, bark, berries, and mixtures thereof. Preferentially, dedifferentiated plant cells are obtained from bark, leaves, buds and fruit skin.

The dedifferentiated plant cells that can be used according to the invention can be obtained from plants obtained by in vivo culture or from culture in vitro. In vivo culture is understood to mean any culture of conventional type, that is to say in soil in the open air or in a greenhouse, or above ground or in a hydroponic medium. By in vitro culture is meant all the techniques known to those skilled in the art which artificially allows the production of a plant or part of a plant. The selection pressure imposed by the physicochemical conditions during the growth of the plant cells in vitro makes it possible to obtain a standardized plant material which is free from contamination and available throughout the year, unlike plants cultivated in vivo. . Preferably, according to the invention, dedifferentiated plant cells originating from in vitro culture are used. The elicitation of the cells is preferably carried out with the cells in a culture medium. The dedifferentiated plant cells that can be used according to the invention can be obtained by any method known from the prior art. In this regard, the methods described by E. F. George and P. D. Sherrington can be cited in Plant Propagation by Tissue Culture, Handbook and Directory of Commercial Laboratories (Exegetics Ltd 1984). The culture media that can be used according to the invention are those generally known to those skilled in the art. Examples of the Gamborg, Murashige and Skoog, Heller, White, etc. environments are found in EF George's Plant Culture Media: Formulations and Uses, DJM Puttock and HJ George (Exegetics 15 Ltd). 1987, volume 1 & 2) complete descriptions of these media. Preferably, the dedifferentiated plant cells cultivated on Murashige and Skoog medium are prepared according to the invention.

Particular features and details of compositions according to the invention will emerge from the following description of exemplary embodiments given by way of example only. Tests demonstrating the effectiveness of the cell mixture of the cosmetic compositions according to the invention will be described hereinafter. In these tests, the following products from plant cells were used: 1. Crushed Centella asiatica Bl cells.

This crushed cell was prepared in the following manner: Step 1: Preparation of dedifferentiated and cultured cells in in vitro culture medium This step of preparing dedifferentiated Centella asiatica cells was carried out in a conventional manner with successive steps of transplanting from stems. The culture medium used is a complete Gamborg B5 medium (G5893) marketed by Sigma-Aldrich (Germany). Other culture media are possible. Step 2: subculture of the dedifferentiated cells of step 1 in an in vitro culture medium for the development and elicitation of the cells.

Elicitation development of the cells of step 1 was carried out in an in vitro medium (for example Gamborg B5) under a gaseous atmosphere containing oxygen and CO2 (CO2 enriched air so that its content is 2%). at 5% by volume) in a cycle comprising 100 periods of low light with a brightness of less than 10 lux (1 to 5 lux) for 1 hour under an atmosphere consisting of moist air (75% relative humidity) enriched of CO2 (so that the CO2 content is 5%) and having a temperature of 30 ° C, these periods of development / elicitation under low (or without) brightness being separated from each other by a period of brightness of more than 100,000 lux for 1 hour under an atmosphere consisting of moist air (relative humidity of 75%) enriched with CO2 (so that the CO2 content of the atmosphere is 5%) and having a temperature of 45 ° vs. The transition from a dark state to an illuminated state is achieved by moving an opaque wall. This culture is operated in a white room.

Step 3: Extraction of dedifferentiated and elicited cells in culture medium in vitro, for example by filtration of the culture medium followed by one or more washing steps, in particular carried out so as not to destroy the structure of the membranes of the cells. cells. Step 4: Drying or lyophilization of the dedifferentiated and elicited cells in culture medium in vitro, this drying operation advantageously being carried out so as not to destroy the structure of the membranes of the cells. This step is advantageously carried out at a temperature below 60 ° C, for example between -60 ° C and 50 ° C.

Step 5: grinding. The grinding is done so that the particle size is less than 5011m. It is advantageous to perform the operation of grinding the dedifferentiated and elicited cells in the presence of one or more agents or excipients of the cosmetic composition so as to ensure the release of the phytoalexins and other compounds from the cells in at least some agents of the cosmetic composition. The grinding was carried out so as to obtain a mean particle size of less than 31.tm (between 1 and 211m). 2. Preparation of an extract of Saussurea involucrata El Dedifferentiated plants of Saussurea involucrata grown in vitro were cleaned. The aerial part of the plants, including flowers, was dried and ground into a powder of less than 50 μm (average particle size by weight). 1 kg of this powder was extracted with 70% ethanol three times under reflux for 2 hours. The alcoholic extract was dried under vacuum until a brownish syrup was obtained. This syrup is then mixed with distilled water to obtain a 5% by weight extract. 3. Preparation of an extract of Panax Pseudoginseng E2 3036962 The same procedure was used for the preparation of the Saussurea involucrata extract from Panax Pseudoginseng plants. 3. Preparation of an extract of Angelica sinensis E3 The same procedure was used for the preparation of the Saussurea involucrata extract from Angelica sinensis plants. 4. Preparation of a Panax Notoginseng Extract (E4) The same procedure was used as for the preparation of the Saussurea involucrata extract from Panax notoginseng plants. Preparation of mixtures to be tested The following mixtures were tested for their effectiveness or properties on the skin (Synthetic reconstituted epidermis): Mixture 1: This mixture M1 was prepared by mixing Centella Asiatica B1 cell grind with vegetable glycerine, the ground material BI representing 20% by weight of the mixture. Mixture 2: This M2 mixture was prepared by mixing vegetable glycerin with Centella Asiatica (B1) cell preseed and Saussurea involucrata (El) extract, the content of B1 in the mixture being 20% by weight, while that in El is 20% by weight. Mixture 3: This M3 mixture was prepared by mixing vegetable glycerin with Centella Asiatica (B1) cell broth, Saussurea involucrata (El) extract, and Panax Pseudoginseng (E2) extract, the B1 content in the mixture being 20% by weight, while that of E1 and E2 is in each case 10% by weight. Mixture 4: This M4 mixture was prepared by mixing vegetable glycerin with Centella Asiatica (B1) cell preseed, Saussurea involucrata (El) extract, Panax Pseudoginseng (E2) extract, Angelina Sinensis extract ( E3) and an extract of Panax notoginseng (E4), the content of B1 in the mixture being 20% by weight, while that of E1, E2, E3 and E4 is in each case 5% by weight.

Blend 5: This M5 blend is placebo and consists only of vegetable glycerin. Mixture 6: This M6 mixture was prepared by mixing vegetable glycerin with the Saussurea involucrata extract (E1) present at 20% by weight in the mixture. Test 1: Evaluation of the effect of mixtures on the energetic and physiological metabolism on SKINETHIC® reconstituted skin (sold by Episkin Lyon France) by determination of the relargarge of CO2 at the level of the epidermis The study was carried out on SKINETHIC® reconstituted epidermis. After placing a quantity of mixture on the reconstituted epidermis, the activity of cell metabolism and the oxygenating power of the metabolism are measured by the determination of the metabolism of D- (14C) -glucose-6-phosphate (288 mCi / mmol, NEN, France) and by the determination of 14CO2. To evaluate the activity of the mixtures, the test was performed under physiological conditions and under asphyxiated conditions. For these asphyxiated conditions, asphyxiation of the epidermis was achieved by oxygen deprivation 24 hours before placing a quantity of mixture on the epidermis. The 14CO2 released from the metabolism of D- (14C) -glucose-6-phosphate was evaluated by capturing 14CO2 on filters placed over the epidermis. D (14 C) -glucose-6-phosphate was contacted with the epidermis throughout the course of the treatment with a mixture for 24 hours. At the end of the treatment time (24 hours), the filters are recovered and the radioactivity of the filters is measured by a scintillation counter.

The amount of mixture applied to an epidermis is expressed in% by weight relative to the weight of the reconstituted epidermis.

Test 1.1 Epidermis treated with mixtures M8 (control), M1 (not the invention), M2, M3 and M4, under physiological conditions applied mixture% by weight of the equilibrium released from the 14CO2 increase mixture% of the release rate with respect to control applied By (cPm) to the weight of the epidermis M5 1.0 1648 +/- 91 - negative control M1 not according to 1.0 1750 +/- 95 +6 the invention M6 not according to 1.0 <1800 < +7 the invention M2 0.5 1995 +/- 117 +21 M2 1.0 2045 +/- 119 +24 M2 2.5 2115 +/- 142 +28 M3 0.5 2119 +/- 150 +29 M3 1.0 2215 +/- 132 +34 M3 2.5 2254 +/- 77 +37 M4 0.5 2253 +/- 87 +37 M4 1.0 2341 +/- 113 +42 M4 2.5 2491 + / Test 1.2 Epidermis treated with mixtures M5 (placebo-control), M1 (not the invention), M2, M3 and M4, under asphyxiated conditions mixture% by weight of measurement of 14CO2 released (cpm) applied growth mixture in% of the release rate applied compared to the ratio to the control condition weight of the epidermis M5 1.0 975 +/- 31 - control condition M1 (not according to the invention) 1.0 1000 +/- 75 +3 M6 not according to the invention 1.0 <+1050 <+8 M2 0.5 1221 +/- 53 +25 M2 1.0 1277 +/- 60 +31 M2 2.5 1350 +/- 77 +38 M3 0.5 1321 +/- 60 +35 M3 1.0 1374 +/- 43 +41 M3 2.5 1410 +/- 82 +45 M4 0.5 1368 +/- 58 +40 M4 1.0 1430 +/- 60 +47 M4 2.5 1500 +/- 89 +54 3036962 13 These tables show a significant and synergistic increase in the amount of CO2 released using mixtures according to the invention, both under physiological conditions and under asphyxiated conditions.

Test 2: Research of the effect of the product on the energetic metabolism of cells in culture. Assay of the cellular adenyl nucleotides (ATP, ADP and AMP) in nmoles / mg of proteins, and calculation of the energy charge (EC) of the cells treated for 5 days with the product.

Human keratinocytes were cultured for 5 days in the presence of a mixture (106 cells per measurement). Once trypsinized, the cells were harvested and the concentrations of the adenyl nucleotides (ATP, ADP and AMD) were determined by HPLC. (ATP: adenosine 5'-triphosphate, ADP: adenosine diphosphate, AMP: Adenosine monophosphate) The EC energy load is calculated by the following formula: CE ([ATP] + 0.5 x [ADP]) / ([ATP ] + [ADP] + [AMP])) The concentrations of ATP, ADP and AMP are expressed in nmoles / mg of protein. This test was carried out on non permeabilized whole cells and with the compositions M5 (control), M1, M6 and M4 (M4 being a mixture giving good results in test 1). The cells were cultured for 5 days with a mixture amount corresponding to 1% by weight of the culture medium for the M5, M1 and M6 mixtures, and at 0.5%, 1% and 2.5% for the M4 mixture. The results are summarized in the table below.

3036962 14 [ATP] Increased Dose [ADP] Growth [AMP] Incrementally [ATP] [ADP] [AMP] (1) (2) (3) Control 1 4700 1620 680 0.79 M5 M1 not 1 4900 +4 1700 +5 700 +3 0.82 invention M6 not 1 4800 +2 1650 +2 690 +2 0.79 invention M4 0.5 6184 +20 1231 +24 796 +24 0.83 M4 1 6848 +33 2195 +42 978 +28 0.79 M4 2.5 7791 +49 2532 +52 1131 +32 0.79 (1) increase [ATP]:% increase over [ATP] obtained with M8 ( 2) increase [ADP]:% increase over [ADP] obtained with M8 5 (3) increase [AMP]:% increase over [AMP] obtained with M8 In the previous table: - ATP concentrations , ADP and AMP are expressed in nmoles / mg proteins (n = 3). 10 - Energy load (CE) = ([ATP] + 1/2 [ADP]) / ([ATP] + [ADP] + [AMP]) This table shows that even if the energy load remains substantially equal, the concentration in Metabolism is increased by the use of the combination of Centella Asiatica cells and Saussurea involucrata extract. Test 3: Evaluation of a mixture according to the invention M4 on the extracellular matrix by determination of total collagen and elastin. Human fibroblasts were cultured for 1 day in the presence of a mixture (106 cells per measurement). After 24 hours of culture, the fibroblasts were recovered by centrifugation. The pellets were digested with collagenases (1 mg / cell pellet) in acetic acid 0.5 ml / l for 24 hours at 4 ° C. After centrifugation at 10,000 g, the collagens were precipitated with 1M sodium chloride (NaCl), the precipitate was resuspended and dialyzed. The primary amino acids were derived by ophthaldehyde acid (OPA) thus eliminating their interference. Hydroxyproline and proline were then derived by NBD-Cl (4-chloro-7-nitrobenzofurazan marketed for example by Sigma-Aldrich) by coupling of the amino groups with NBD-Cl. The NBD-Hyp was separated and identified by reverse phase HPLC. For the development of the separation of amino acid derivatives, a coupling to NBD-C1 of a standard containing hydroxyproline was first performed. The hydroxyproline was assayed by fluorescence measurement after separation by reverse phase HPLC, for this assay was used: Automatic Injector Column Ultrasep C18 (30 cm x 0.18 cm), 6 lm of porosity Fluorescence detector The The mobile phase consisted of a mixture of acetonitrile / 0.1 mol / l sodium phosphate buffer, pH 7.2 (9:91 v / v), the flow rate was set at 1 ml / min, elution was was performed in static mode and the cycle was 10 minutes. The mobile phase was previously filtered and degassed before use. The solutions were prepared as follows: NBD-Cl = 25 mmol dissolved in methanol, 30-OPA = 150 mmol / l dissolved in methanol, phosphate buffer = 0.4 mmol / l, pH adjusted to 7.2 . The standard range was prepared from a solution of hydroxyproline at 50 mg / l. Successive dilutions make it possible to obtain solutions ranging from 0.5 to 40 mg / l. Derivatization and establishment of the standard curve were performed from 101 μl of a standard solution at different concentrations mixed with 10 μl of the buffer. After addition of 50 OPA and stirring, the tubes were kept for 5 minutes at room temperature and 10111 of the NBD-CI solution were added. The derivate is at 60 ° C in a water bath for 3 minutes, protected from light. The tubes were then removed and the orange color was used to check the derivatization. They were then put in the ice to ensure cooling. 50% of this mixture was then injected into the column in order to obtain the calibration curve which must be linear. For the determination of elastin, the treatment of the cells with the product under study was carried out as previously described for the determination of collagens. The elastin assay was performed according to the protocol indicated in the elastin assay kit (FASTIN TM, Diodye Science). The following table gives the hydroxyproline concentration (in mg / l) and the elastin content of the elastin. proteins for keratocytes contacted with 1% of the M8 mixture (control), 0.5%, 1 and 2.5% of the M4 mixture, the percentage being the percentage by weight relative to the culture medium. The increase is given in% in relation to the control. Mixture Amount increase concentration Elastin increase% hydroxyproline% lag / g% mg / 1 protein M5 1 3.45 15.9 M4 0.5 4.45 +29 20.7 +30 M4 1.0 4.61 +34 22.4 +41 M4 2.5 4.76 +38 23.2 +46 3036962 17 With a mixture according to the invention, we obtain a significant increase, both in concentration of hydroxyproline and elastin in the proteins.

Test 4: evaluation of the proteasomic activity with a mixture according to the invention.

In a first step, three proteasome subunits were isolated and compared from normal senescent cells (from a healthy 62-year-old volunteer donor). The cell extracts were centrifuged at 10,000 xg for 16h at 4 ° C. The pellet was dissolved in Tris-HCl buffer (25 mM, pH 7.5) and then plated on a CNBr Sepharose column (containing a monoclonal antibody directed against the subunit of the human proteasome to be purified) previously equilibrated with the buffer (25 mM Tris-HCl, pH 7.5). The column was then washed with the same buffer and the proteasome was eluted with Tris-HCl containing 2M NaCl (pH 8) and dialyzed for 16 h at 4 ° C (or deposited on a gel filtration column ( PD10 Sephadex) A purified proteasome sample was mixed with denaturing loading buffer (0.1% SDS) and incubated at 100 ° C. for 5 min The proteins were separated by electrophoresis in a 12% acrylamide gel % and (SDS-PAGE) The migration was carried out at room temperature at a constant voltage of 80V for 30 min, then 120V for 2h In a second step, the activity of the three subunits of the proteasome was evaluated under normal physiological conditions (normal senescent cells).

The peptidase activities of the proteasorne have been measured by the use of synthetic peptides whose N-terminal ends are blocked and the C-terminal ends are linked by an isopeptide bond to a fluorescent radical: 7-amido-4-methyl coumarin (MCA). These non-fluorescent radicals, when bound to the peptides, become fluorescent in the free state after proteolytic cleavage. The mixture, containing either 50 μg of crude protein total protein or 31 μg of purified proteasome (in 25 mM Tris-HCl pH 7.5) is incubated at 37 ° C with the substrate in a final volume of 200 μl for 30 minutes. minutes. The reaction is stopped by 3000 of acid or ethanol. After addition of 2 ml of distilled water, the fluorescence is measured using a microplate reader at 350/440 nm excitation and emission wavelengths for the MCAs. The proteasome activities are determined as the difference between the total activity and the remaining activity of the crude extract in the presence of 20 μM proteasome inhibitor. Oxidized Proteins The detection of the oxidized proteins was carried out using the Oxyblot kit. The cytosolic extracts of the epidermis (treated or not) are treated for 15 min. by 2,4-dinitrophenylhydrazine (to derivatize the carbonyl groups of the proteins), then separated by 12% acrylamide gel electrophoresis (SDS-PAGE) at the rate of 10 μg of protein per well. They are then transferred to Nitrocellulose Hybond membrane. The hydrazones formed are immunodetected with polyclonal rabbit antibodies against the 2,4 dinitrophenyl radical (Sigma, ref D-9656). In order to detect ubiquitinated or modified proteins by the addition of 4-hydroxy-2-nonenal, 20 μg of proteins are deposited on 12% polyacrylamide gel and the Western blots are revealed using polyclonal antibodies directed against ubiquitin. Detection of antigen-antibody complexes is performed with rabbit secondary antibodies coupled to peroxidase.

The following table gives the results of the tests with the mixture M5 (control used at 1% by weight of the culture medium), and with the mixture M4 (at the rate of 0.5, 1.0 and 2, 5% by weight of the culture medium). Mixture% P2OS P2OS P26S P26S PA28 PA28 P Ox P Ox%%%% M5 1 7.56 60.2 176.8 460 M4 0.5 9.58 27 74.1 23 210.4 19 345 -25 M4 1, 0 10.25 36 80.3 33 220.7 25 321 -30 M4 2.5 11.23 49 86.4 44 265.0 50 280 -39 5 P2OS: 20S proteasome in nmole / minute / mg P20S%: increase in% relative to the P2OS of the P26S control: Proteasome 26S in nmol / minute / mg P26S%: increase in% relative to the P26 of the control PA28: proteasome complex PA28 (nmol / min / mg) PA28%: increase in% by compared to the PA28 control P Ox: density of oxidized protein and ubiquitinated (U / mm2) P Ox%: increase in% relative to the density of protein control This table shows that with the invention, it is possible to reduce the density of oxidized and ubiquitinated protein, but increase the proteasomic activity. Test 5: Tolerance test An in vitro dermal and ocular tolerance study was carried out on a preliminary basis for the preparation of cosmetic preparations. These tests revealed a perfect local tolerance (cutaneous and ocular) of the M4 mixture at a concentration of 0.3%.

The mixtures M2 to M6 or the ground and extracted can be directly used to form particular cosmetic compositions for topical use. Some examples of non-limiting examples of such compositions for topical use are given below. EXAMPLE 1 Dispersion of Crushed B1 Proteins with Extract E and Advantageously with E2 and / or E3 Extract in a Cosmetic Base The ground material B1 is dispersed in the following base, with addition of extract E1 and advantageously of extract E2 and / or E3: - Deionized water 85, 41% Mineral oil 9.00% Cetyl alcohol 3.00% ceteareth - 20 0, 75% 15- ground (at least B1) 0, 20% Extract (at least El) 0.20% fragrance 0, 15% carbomer 0, 10% methylchloroisothiazoline 20 and methylisothiazoline [kathon CG] 0, 065% - hydroxide sodium hydroxide (45%) 0.06% - butylated hydroxyanisole 0.06% TOTAL 100.00% The composition obtained shows a homogeneous dispersion of the cells in the cream and a very fine particle size distribution. The property study showed the absence of germs and fungi as well as a remarkable stability of the composition.

EXAMPLE 2 Dispersion of Crushed Cell in a Cosmetic Base The ground material B1 is dispersed in the following base, with the addition of extract E1 and advantageously of extract E2 and / or E3: water 46.59% laureth sodium sulfate (25%) 36, 40% - PEG-7 glyceryl cocoate 2.00% - laureth-2 1, 50% 10-laureth-11 sodium carboxylate 4.00% cocamidopropyl betaine & benzoic acid 3.48% sodium chloride 1, 60% - propylene glycol 1.00% - perfume 0, 13% 15- PEG-40 hydrogenated beaver oil & propylene glycol & water 0, 50% oleth-10 0, 50% sodium phosphate 0, 30% disodium phosphate 0.88% 20-citric acid (50%) 0.52% sodium benzoate 0.50% Broyate Blet extract El (plus possibly E2 and / or E3) 0.50% salicylic acid 0.25% phenoxyethanol 0.20% TOTAL 100.00% The composition obtained shows a homogeneous dispersion of the cells (advantageously ground) and extracted (s) in the cream, and a very fine particle size. The property study showed the absence of germs and fungi as well as remarkable stability of the composition.

EXAMPLE 3 Creams-aqueous phase A: demineralized water combined with a moisturizing product-oily phase B: emulsifier + emollient or emollient material + oil 5 -phase C: preservative, perfume -phase D: active product: crushed B1 cells with extract El and with or without extract E2, E3, in the form of a viscous suspension or a gel or a substantially dry powder.

Example 4: Lotions Lotions containing only an aqueous phase A: demineralized water, propylene glycol, preservative, perfume; and active product: ground product and extract (s), in the form of a viscous suspension or a substantially dry gel or powder. EXAMPLE 5 Shampoos Shampoos containing only a demineralized water aqueous phase A, detergents, foaming agents, thickeners, perfume and active product: ground product and extract (s), in the form of a viscous suspension or a gel or a substantially dry powder. EXAMPLE 6 Gels 25 - Hydrogels and oleogels obtained by adding to the aqueous phase A or to the oily phase B emulsifying and thickening agents are considered. step C: perfume, preservative -phase D: ground product and extract (s), in the form of a viscous suspension or a gel or a substantially dry powder.

EXAMPLE 7 Solutions Solutions containing only an aqueous phase A essentially based on demineralised water, perfume, preservative and active product: ground product and extract (s), in the form of a viscous suspension or a gel or a substantially dry powder. EXAMPLE 8 Milks 10-aqueous phase A: essentially based on deionized water-oily phase B: oil + emulsifier + emollient -phase C: preservative + moisturizing product -phase D: active product: ground product and extract (s), under form of a viscous suspension or a gel or a substantially dry powder.

In the examples given above, concerning creams, gels or milks, the different phases A, B, C and D, in proportions which may vary from one another according to the desired applications, are mixed in the usual way, as the usually performs the skilled in this field.

With regard to lotions, solutions and shampoos, the composition for topical use contains the various constituents, the contents of which can be varied according to the applications, mixed in the single aqueous phase, as is usually carried out by those skilled in the art in this field. field.

The proportion of each ground material and extract (s) depends on the nature of the topical composition and the desired application. It is advantageously between 0.01 and 5%, but can reach up to 25%.

Obviously, the invention is not limited to the embodiments given above and it is possible to make the composition for topical use in other forms, such as oil, ointment, lacquers, blushes (bottom of complexion, powder, lipstick, pencil, mascara or cosmetic product to highlight the eyes by coloring the eyelashes and / or giving them more length or apparent thickness, eye shadow) which also fall within the scope of the invention. 5

Claims (12)

REVENDICATIONS1. Cosmetic composition containing at least (a) Centella Asiatica cells or a crushed material of such cells, (b) an extract of Saussurea involucrata, and (c) a carrier for cosmetic application. 2. Composition according to claim 1, characterized in that it contains at least (a) dedifferentiated and eluted Centella Asiatica cells, preferably in the form of ground matter, and (b) an extract of Saussurea involucrata. 3. Composition according to claim 1, characterized in that it contains at least (a) a crushed dedifferentiated and eluted Centella Asiatica cells, and (b) an extract of Saussurea involucrata. 4. Composition according to any one of the preceding claims, characterized in that it further contains an extract of plants of the family consisting of Araliaceae and Apiaceae, said extract being cosmetically acceptable. 5. Composition according to the preceding claim, characterized in that it also contains an extract of plants of the genus Panax of the family of araliaceae and / or of the genus Angelica or Angelina of the family Apiaceae. 25 6. Composition according to claim 5, characterized in that it contains an extract of Panax Pseudoginseng, Panax ginseng, Panax Notoginseng, Angelica or Angelina Sinensis, and a mixture of such extracts. 7. A composition according to any one of the preceding claims, characterized in that it contains an effective amount of Centella Asiatica cells or crushed such cells, and Saussurea involucrata extract for its use. to ensure an energizing activity for the skin and / or proteasomic activity of the skin. 8. Composition according to one of the preceding claims, characterized in that it contains from 0.02% to 10% by weight, preferably 0.05 to 0.5% by weight, in particular 0.06% by weight. % to 0.2% by weight of Centella Asiatica cells or crushed such cells, and from 0.02% to 10% by weight, preferably from 0.05 to 0.5% by weight, in particular from 0 to From 06% to 0.2% by weight of extracts of Saussurea involucrata. 9. Composition according to any one of the preceding claims, characterized in that it contains a crushed dedifferentiated and eluted Centella Asiatica cell having a weight average size of less than 101.1m, in particular less than 5gm, preferably less than liam. 10. Composition according to any one of the preceding claims, adapted for cleaning the skin. 11. Composition according to any one of the preceding claims, characterized in that it is in the form of powder, creams, pastes, gels, suspensions, fluid or liquid compositions, solid or semi-solid compositions. , stick, sticks, pencil, solid gel or semi-solid release 12. Composition according to any one of the preceding claims, characterized in that it contains an effective amount of Centella Asiatica cells or a crushed such cells, and Saussurea involucrata extract, for its use to induce a stimulation of the basal cell respiration rate of non-permeabilized human keratinocytes of at least 5%, advantageously at least 20%, preferably at least 50%, in particular at least 75%, and / or 3036962 27 for its use for inducing a stimulation of the mitochondrial respiration rate of permeabilized human keratinocytes by at least 5%, advantageously by at least 20%, preferably by at least 50%, in particular by at least 75%, presence of pyruvate-malate, and / or for its use to induce a stimulation of the synthesis rate of adenisone triphosphate (ATP) of non-permeabilized human keratinocytes by at least 0.5%, advantageously by at least 10% , d Preferably at least 20%, in particular at least 50%, more preferably at least 75%, and / or for use in inducing stimulation of the synthesis rate of Adenisone Triphosphate (ATP). mitochondrial membrane of permeabilized human keratinocytes of at least 0.5%, advantageously at least 10%, preferably at least 20%, in particular at least 50%, in the presence of pyruvate-malate, and / or for its use in inducing sum stimulation (cellular Adenisone Triphosphate (ATP) synthesis rate + synthetic Adenosine monophosphate (AMP) synthesis rate + Adenosine Diphosphate Cell Synthesis (ADP) rate ) at least 10%, advantageously at least 20%, preferably at least 50%, in particular at least 100% of non-permeabilized human keratinocytes after 5 days of contact, while advantageously maintaining a ratio of the energetic load between 0, 74 and 0, 85, and / or for its use for p to allow differentiation and proliferation of human keratinocytes in culture, and / or for its use to allow a better cohesion of the cells of the keratinocytes of human skin, and / or for its use to ensure a vital effect of force on the skin, and / or for its cosmetic use to ensure a moisturizing effect by NMF, and / or for its use to ensure a homeostasis effect, and / or for its use to ensure cellular cohesion of keratinocytes, and / or 3036962 28 for its use to ensure a cellular renewal effect by proliferation and differentiation of cutaneous cells. 5