NOVEL COMPOSITION BASED ON POLYCAFFEOYLQUINIC ACIDS, COSMETIC USE THEREOF AND COSMETIC COMPOSITIONS COMPRISING SAME

Abstract

Disclosed is a composition including, per 100% of its mass: a) 60.0% to 75.0% by mass of an organic solvent (OS.sub.1) chosen from 1,2-propanediol, 1,3-propanediol, 1,4-butanediol, 1,3-butanediol, 1,2-butanediol, 2-methyl-2,4-pentanediol, 1,6-hexanediol, 1,8-octanediol, or a mixture of these compounds; b) 0.1% to 2.0% by mass of a composition (ES) including an amount by mass x.sub.1, expressed as mass equivalent of 1-O-(2-caffeoyl)maloyl-3,5-di-O-catfeoylquinic acid, of greater than or equal to 200 mg/g of a compound of general formula (I):

##STR00001##

in which Q.sub.1, Q.sub.2, Q.sub.3, Q.sub.4 and Q.sub.5 represent, independently of each other, a hydroxyl radical or a salt thereof or a radical chosen from caffeoyl, maloyl, caffeoyl maloyl and maloyl caffeoyl radicals, it being understood that at least one of these radicals Q.sub.1, Q.sub.2, Q.sub.3, Q.sub.4 and Q.sub.5 represents neither the —OH radical nor a salt thereof; and c) 20.0% to 35.0% by mass of water. Related treatments are also disclosed.

Claims

1. A process for preparing a composition (C.sub.1) comprising the following successive steps: a) cultivation under soilless conditions of the plant Arctium lappa fed with a nutrient solution, so as to obtain a biomass (BM.sub.1); b) immersion of the roots of said biomass (BM.sub.1) obtained in the preceding step a) in a medium (S.sub.1), such that the biomass (BM.sub.1)/mixture (S.sub.1) ratio is between 0.5 kg/l and 1.5 kg/l, said medium (S.sub.1) comprising, per 100% of its own mass, from 20% to 35% by mass of water, the pH of which has been adjusted to a value of between 1.5 and 3.5 by addition of a protic acid chosen from sulfuric acid, phosphoric acid and hydrochloric acid, and from 65% to 80% by mass of an organic solvent (OS.sub.1) chosen from 1,2-propanediol, 1,3-propanediol, 1,4-butanediol, 1,3-butanediol, 1,2-butanediol, 2-methyl-2,4-pentanediol, 1,6-hexanediol, 1,8-octanediol or a mixture of these diols; c) separation of the roots of the biomass on conclusion of the treatment defined in step b), in order to isolate a liquid phase (L.sub.1); d) immersion of the roots of said biomass (BM.sub.2) resulting from step c) in said medium (S.sub.1); in a biomass (BM.sub.2)/mixture (S.sub.1) ratio of between 0.1 kg/l and 1.5 kg/l; e) separation of said biomass (BM.sub.2) on conclusion of the treatment defined in step d), in order to isolate a liquid phase (L.sub.2); f) filtration of said liquid phase (L.sub.3) obtained in step d), in order to isolate a liquid phase (L.sub.3), and g) mixing said liquid phases (L.sub.1) and (L.sub.3), then, if necessary, of addition of water and/or of said organic solvent (OS.sub.1), so as to obtain the composition (C.sub.1).

2. The process of claim 1 wherein the composition (C.sub.1) comprises, per 100% of mass: a)- from 60.0% by mass to 75.0% by mass of an organic solvent (OS.sub.1) chosen from 1,2-propanediol, 1,3-propanediol, 1,4-butanediol, 1,3-butanediol, 1,2-butanediol, 2-methyl-2,4-pentanediol, 1,6-hexanediol, 1,8-octanediol, or a mixture of these compounds; b)- from 0.1% by mass to 2.0% by mass of a composition (ES) comprising an amount by mass x.sub.1, expressed as mass equivalent of 1-O-(2-caffeoyl)maloyl-3,5-di-O-caffeoylquinic acid, of greater than or equal to 200 mg/g of at least one compound of general formula (I): ##STR00007## in which Q.sub.1, Q.sub.2, Q.sub.3, Q.sub.4 and Q.sub.5 represent, independently of each other, a hydroxyl radical or a salt thereof or a radical chosen from: (i)- the caffeoyl radical of formula (II): ##STR00008## (ii)- the maloyl radical of formula (IIIa) or (Ib): ##STR00009## (iii)- the caffeoyl maloyl radical of formula (IVa) or (Ib): ##STR00010## (iv)- the maloyl caffeoyl radical of formula (Va), (Vb), (Vc) or (Vd), ##STR00011## it being understood that at least one of these radicals Q.sub.1, Q.sub.2, Q.sub.3, Q.sub.4 and Q.sub.5 represents neither the —OH radical nor a salt thereof; and c)- from 20.0% by mass to 35.0% by mass of water.

3. The process of claim 2 wherein said composition (ES) comprises: at least one compound of formula (Ia) corresponding to formula (I) for which Q.sub.1 represents the maloyl radical of formula (IIIa) or of formula (IIIb) and Q.sub.3 and Q.sub.4 and Q.sub.5, which are identical, each represent the caffeoyl radical of formula (II); a compound of formula (Ib) corresponding to formula (I) for which Q.sub.1 represents the caffeoylmaloyl radical of formula (IVa) or of formula (IVb), Q.sub.3 and Q.sub.5 each represent the caffeoyl radical of formula (II) and Q.sub.4 represents a hydroxyl radical, and at least one compound of formula (Ic) chosen from: the compound of formula (I.sub.c1) corresponding to formula (I) for which Q.sub.1 and Q.sub.5 each represent the caffeoyl radical of formula (II), Q.sub.3 represents a hydroxyl radical and Q.sub.4 represents the caffeoylmaloyl radical of formula (IVa) or of formula (IVb); and the compound of formula (I.sub.c2) corresponding to formula (I) for which Q.sub.1 and Q.sub.4 represent the caffeoyl radical of formula (II), Q.sub.3 represents a hydroxyl radical and Q.sub.5 represents the caffeoylmaloyl radical of formula (IVa) or of formula (IVb).

4. The process of claim 2 wherein the organic solvent (OS.sub.1) is 1,2-propanediol.

Description

DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0148] The examples that follow illustrate the invention without, however, limiting it.

A.sub.1) -Preparation of a Composition (C.sub.1A) According to the Invention

[0149] Burdock or Arctium lappa plants were obtained beforehand by germination of seeds over a period of 60 days under standard conditions, so as to reach a size of about 10 to 15 cm, then they are removed from their pots and placed under aeroponic culture conditions, consisting in leaving the burdock plants to soak in a nutrient solution characterized by an electrical conductance of between 1.0 mS (millisiemens) and 1.2 mS and by an N/P/K (nitrogen/phosphorus/potassium) mass ratio provided by the fertilizer of about 15/10/30; for 6 weeks at a temperature regulated at 20° C. A root yield of 754 g per square meter is thus obtained.

[0150] The fresh roots of the biomass thus obtained are collected and immersed for 15 minutes at 25° C., at a rate of 1.0 kg of biomass per liter, in a bath comprising, per 100% of its mass, 70% by mass of 1,2-propanediol and 30% by mass of distilled water, the pH of which was adjusted beforehand to 2.0±0.2, by adding a solution containing 75% by mass of phosphoric acid.

[0151] The plants are then removed from their exudation bath (L.sub.1), the roots are drained and then chopped up, and the remaining biomass is then macerated again for 48 hours at 25° C., at a rate of 0.5 kg of biomass per liter, in a new bath comprising a mixture comprising, per 100% of its mass, 70% by mass of 1,2-propanediol and 30% by mass of distilled water, the pH of which was adjusted beforehand to 2.0±0.2, by adding a solution containing 75% by mass of phosphoric acid.

[0152] On conclusion of this maceration phase, the biomass is separated from the maceration liquid (L.sub.2) and said liquid (L.sub.2) is filtered with a 50 micrometer filter bag.

[0153] The liquids (L.sub.1) and (L.sub.2) are pooled and the mixture obtained is then supplemented with 1,2-propanediol to adjust its mass content to 70%. The liquid (La) thus obtained is filtered through a 1 micrometer membrane so as to clarify it, and then through a 0.2 micrometer membrane to obtain the expected composition (C.sub.1A).

A.sub.2) -Example of Preparation of a Comparative Composition (C.sub.comp)

[0154] The plantlets resulting from the same batch of seeds as that used to obtain the plantlets subsequently cultivated aeroponically (example A.sub.1) are used for cultivation of said plantlets in soil for a period of time of six weeks.

[0155] At the end of this period, the plants are removed from the pots, the fresh roots are cleaned, cut up and ground, and said ground material obtained is then extracted according to a conventional liquid/solid extraction process (maceration, agitation, filtration) using a 70/30 1,2-propanediol/distilled water solvent mixture, at a temperature of 25° C., with a fresh roots/solvent medium mass ratio of 0.5 kg of biomass per 1 liter of 1,2-propanediol and water mixture; the value of the pH of the distilled water having been adjusted beforehand to 2.0±0.2 by addition of a solution containing 75% by mass of phosphoric acid.

[0156] At the end of this extraction phase, the biomass is separated from the liquid, which is subsequently filtered with a 50 micrometer filter bag, then with a 1 micrometer membrane, so as to clarify it, and finally under sterilizing filtration with a 0.2 micrometer membrane, so as to achieve the composition (C.sub.comp).

[0157] A.sub.3) Analytical Characterization of the Composition (C.sub.1A) According to the Invention and of the Comparative Composition (C.sub.comp)

[0158] The characteristics of compositions (C.sub.1) (C.sub.comp) are collated in table 11 below:

TABLE-US-00007 TABLE 11 Composition Characteristics Analytical method (C.sub.1A) (C.sub.comp) Appearance Visual Yellow Yellow Liquid Liquid 1,2-Propanediol content GC (headspace) 71.4% 70.0% pH According to the 3.3 3.2 standard NFT 73-206 Solids content as % Oven at 105° C., 0.21% 1.12% by mass 12 hours Water % by mass According to the 28.39% 28.88% standard NFT 73201 Total content of compounds UPLC-MS* 613.3 169.5 of formulae (Ia), (Ib), (Ic.sub.1) and (Ic.sub.2) (mg/gram of solids content) *UPLC-MS: Machine: Shimadzu Nexera X2; Column: Waters Xterra RP C18; 250 × 4.6 mm; mobile phase (with gradient): water + formic acid/acetonitrile; UV detector (330 nm)
B) Demonstration of the Moisturizing Properties of Composition (C.sub.1A) According to the Invention and of the Comparative Composition (C.sub.comp).

B.SUB.1.) Demonstration of the Protection Against Impairment of the Skin's Barrier Function, on Samples of Reconstructed Human Epidermis.

B1.1. Principle of the Method

[0159] The strains Staphylococcus epidermidis and Staphylococcus aureus were cultured in BHI(Brain Heart Infusion) and NB(Nutrient Broth) medium, respectively, at 37° C. for 24 hours. Samples of reconstructed human epidermis with a surface area of 0.5 cm.sup.2, cultivated at 37° C. and under 5% CO.sub.2, were colonized first with the strain Staphylococcus epidermidis for a period of 6 hours, and were then colonized with the strain Staphylococcus aureus for 24 hours.
Composition (C.sub.1A) according to the invention (1% v/v) was added to the samples of reconstructed human epidermis at the same time as the strain Staphylococcus epidermidis and then again with the strain Staphylococcus aureus.
The barrier function of the samples of reconstructed human epidermis thus treated was evaluated:

[0160] by measurement of the transepithelial electrical resistance (TEER) of the samples of reconstructed human epidermis and

[0161] by histological evaluation of said samples of reconstructed human epidermis, and more particularly by staining with hematoxylin and eosin, and by a “score” of said staining.

[0162] In the context of the histological evaluation, the effects on the barrier function of the samples of reconstructed human epidermis were evaluated by a histological score on staining with hematoxylin and eosin, which was attributed as follows:

[0163] 0=standard: no significant modification of the reference morphology

[0164] 1=mild: significant modification of the stratum corneum

[0165] 2=moderate: significant modifications of the stratum corneum and of the granular layer, decrease in keratohyalin and a few necrotic cells

[0166] 3=strong: significant modifications in the basal layer with necrotic cells and intercellular holes and edemas

[0167] 4=severe: loss of intercellular connection, detachment of the tissue from the polycarbonate filter, necrotic cells, absence of specific labeling.

[0168] A product is judged to be protective on the barrier function of human epidermis if the histological score is graded “standard” (score 0) or “mild” (score 1).

[0169] The equilibrium of the microbiota of the reconstructed human epidermis, with or without application of composition (C.sub.1A), was evaluated by studying the formation of ultrastructures such as colonies and biofilms by SEM.

B.1.2. Results

[0170] B.1.2.1 Results obtained on the protection of the barrier function of the samples of reconstructed human epidermis by measuring the transepithelial electrical resistance (TEER)

[0171] The TEER measurements performed on the samples of reconstructed human epidermis, as a function of the associated treatments, are collated in Table 7. A decrease in the transepithelial electrical resistance (TEER) is evidence of degradation of the barrier function of the epidermis and consequently constitutes one of the factors for skin dehydration and of the unesthetic effects that this dehydration may cause. The difference A between the transepithelial electrical resistance of the surface of the reconstructed human epidermis after colonization and before colonization is also calculated. The percentage of protection is also calculated according to the formula: % Protection=R.sub.[reconstructed human epidermis colonized with Staphylococcus epidermidis+Staphylococcus aureus and treated with (C1A) 1% (v/v)]- R.sub.[reconstructed human epidermis colonized with Staphylococcus epidermidis+Staphylococcus aureus without addition of (C1A)]/R.sub.[untreated reconstructed human epidermis (control)]−R.sub.[reconstructed human epidermis colonized with Staphylococcus epidermidis+Staphylococcus aureus without addition of (C1A)]

TABLE-US-00008 TABLE 7 Untreated reconstructed human epidermis (control) At t = 0 Electrical resistance (in Ohm .Math. cm.sup.2) R.sub.0 = 8073.33 ± 1913.60 At t = 30 hours Electrical resistance (in Ohm .Math. cm.sup.2) R′.sub.0 = 6900.00 ± 1489.80 Δ.sub.0 = R′.sub.0 − R.sub.0 −1173.33 Reconstructed human epidermis colonized with Staphylococcus epidermidis Staphylococcus aureus without addition of Composition (C.sub.1A) Before colonization (t = 0) Electrical resistance (in Ohm .Math. cm.sup.2) R.sub.1 = 8094.44 ± 1486.73 After colonization (t = 30 hours) Electrical resistance (in Ohm .Math. cm.sup.2) R′.sub.1 = 4726.11 ± 2067.16 Δ.sub.1 = R′.sub.1 − R.sub.1 −3368.33 (100/Δ.sub.0) × (Δ.sub.1 − Δ.sub.0) 187%  Reconstructed human epidermis colonized with Staphylococcus epidermidis + Staphylococcus aureus and treated with Composition (C.sub.1A) 1% (v/v) (0.0136% of dry extract of composition (C.sub.1A)) Before colonization Electrical resistance (in Ohm .Math. cm.sup.2) R.sub.2 = 7647.22 ± 1566.61 After colonization Electrical resistance (in Ohm .Math. cm.sup.2) R′.sub.2 = 5645.56 ± 911.52 Δ.sub.2 = R′.sub.2 − R.sub.2 −2001.66 (100/Δ.sub.0) × (Δ.sub.2 − Δ.sub.0) 71% % of protection [(R′.sub.2 − R′.sub.1)/(R′.sub.0 − R′.sub.1)] 42%

[0172] When the samples of reconstructed human epidermis are colonized with Staphylococcus epidermidis and Staphylococcus aureus, the difference in TEER measured before and after the start of said colonization is −3368.33 Ohm.Math.cm.sup.2, and shows an increase of 187% relative to the samples of non-colonized and untreated reconstructed human epidermis (−1173.33 Ohm.Math.cm.sup.2).

[0173] When the samples of reconstructed human epidermis are placed in contact with composition (C.sub.1A) at the same time as their colonization with Staphylococcus epidermidis and Staphylococcus aureus, the difference between the TEER values measured before and after said colonization is −2001.66 Ohm.Math.cm.sup.2, which represents an increase of 71% relative to the samples of non-colonized and untreated reconstructed human epidermis (−1173.33 Ohm.Math.cm.sup.2) and a protection of 42% relative to the samples of reconstructed human epidermis colonized with Staphylococcus epidermidis and Staphylococcus aureus.

[0174] As a result, application to the skin of a composition comprising composition (C.sub.1A) makes it possible to prevent the degradation of the barrier function of the epidermis of the skin, before the skin is subjected to the action of bacteria known for their degrading effects on the barrier function of said epidermis of the skin.

[0175] B.1.2.2 Results obtained on the protection of the barrier function of samples of reconstructed human epidermis by histological evaluation of said samples of reconstructed human epidermis, by staining with hematoxylin and eosin.

[0176] The staining with hematoxylin and eosin of the epidermal samples was evaluated by attributing a “score” as described previously, and the results are collated in table 8 below:

TABLE-US-00009 TABLE 8 Histological score Untreated reconstructed human epidermis (control) 0: no significant modification vs the reference morphology Human epidermis colonized with Staphylococcus epidermidis + Staphylococcus aureus without addition of Composition (C.sub.1A) 2: modification of the structure of the viable epidermis with more intercellular spaces and regrouping of cells. Reconstructed human epidermis colonized with Staphylococcus epidermidis + Staphylococcus aureus and treated with Composition (C.sub.1A) 1% (v/v) (0.0136% of dry extract due to the plant of composition (C.sub.1A)) 1: reduction of the damage, in particular at the basal level and in the stratum corneum where a more compact lamellar structure is observed

[0177] When the samples of reconstructed human epidermis were placed in contact with Composition (C.sub.1A) at the same time as their colonization with Staphylococcus epidermidis and Staphylococcus aureus, the histological score was evaluated at a level of 1, which is evidence of a reduction of the damage, in particular at the basal level and in the stratum corneum where a more compact lamellar structure is observed.

[0178] As a result, the application to the skin of a composition comprising Composition (C.sub.1A) makes it possible to prevent degradation of the tissue cohesion and consequently of the barrier function of the epidermis in the face of an invasion of the transient flora. Furthermore, the profile of colonization with the two bacteria (Staphylococcus epidermidis and Staphylococcus aureus) was evaluated by scanning electron microscopy (SEM, Zeiss Sigma Electron Microscope).

[0179] When the samples of reconstructed human epidermis were colonized only with Staphylococcus epidermidis, the bacterium is present homogeneously on the surface of the reconstructed human epidermis, forming large aggregates and developing a biofilm characterized by filamentous polysaccharide structures observed at a magnification of ×10 000 of the electron microscope.

[0180] When the samples of reconstructed human epidermis were colonized with Staphylococcus epidermidis and Staphylococcus aureus, the appearance of several spherical aggregates of Staphylococcus aureus on the surface of the reconstructed human epidermis is observed, whereas a film of Staphylococcus epidermidis remains visible on the surface of said epidermis.

[0181] The presence of large aggregates of Staphylococcus aureus is also observed, with a magnification of ×10 000 of the electron microscope, forming a three-dimensional structure, thus indicating an early stage of development of a biofilm at the surface of the epidermis.

[0182] After application to the skin of a composition comprising Composition (C.sub.1A), the spherical aggregates of Staphylococcus aureus are no longer present, thus meaning that the addition of Composition (C.sub.1A) makes it possible to prevent the adhesion of the bacteria to the surface of the epidermis and the formation of the Staphylococcus aureus biofilm. The Staphylococcus epidermidis biofilm still remains visible on the surface of the epidermis.

[0183] These observations show that Composition (C.sub.1A) makes it possible to reduce the adhesion and thus the formation of biofilms of pathogenic opportunistic bacteria, for instance Staphylococcus aureus, without impairing the presence of commensal bacteria such as Staphylococcus epidermidis.

B.1.3. Conclusions

[0184] The combination of measurement of the transepithelial electrical resistance and of the histological evaluation of samples of reconstructed human epidermis, before colonization with a commensal bacterium of the cutaneous flora, and then with a pathogenic bacterium, constitutes a model for studying the impairment of the barrier function and the equilibrium of the microbiota of said epidermis, and the incidence of prior treatments with compositions or extracts or complex formulations.

[0185] The set of results and observations collected in sections B.1.2.1 and B.1.2.2 shows that composition (C.sub.1A) makes it possible to prevent degradation of the barrier function of the epidermis of human skin and consequently to prevent dehydration of the epidermis of the human skin in a model in which it is colonized with a commensal bacterium of the cutaneous flora, and then with a pathogenic bacterium, and the unesthetic effects associated therewith, for example dry, rough skin, which may be accompanied by inflammation and itching when it is associated with the presence of pathogenic bacteria.

B.SUB.2.) Demonstration of the Improvement in the Barrier Function of the Skin by Increasing the Keratinocyte Differentiation

B.2.1 Principle of the Method

[0186] The evolution of differentiation of the epidermal keratinocytes constitutes a means for showing the effect of a treatment of said epidermis on the evolution of the barrier function of the epidermis of human skin, and consequently on the evolution of the state of moisturization of the epidermis of human skin.

[0187] To do this, normal human keratinocytes were cultured at 37° C. and 5% CO.sub.2. After a period of 4 days, they were treated with the products whose action it is desired to evaluate.

[0188] After culturing for a further 72 hours at 37° C. and 5% CO.sub.2 with the test products, the cell lawns were rinsed, dried and then fixed with glacial methanol. Filaggrin, a keratinocyte differentiation marker, was detected and quantified by fluorescent immunolabeling with an anti-filaggrin primary antibody and a fluorescent secondary antibody to reveal the former. The nuclei were detected and quantified by marking with Hoechst, a fluorescent stain which binds to DNA and thus labels the nuclei. This last labeling was performed to normalize the results obtained previously on filaggrin.

B.2.2 Results

[0189] The filaggrin measurements taken on keratinocytes cultured under different conditions, and notably in the presence of Composition (C.sub.1A) according to the invention and in the presence of the comparative composition (C.sub.1Com), are collated in Table 9.

[0190] An increase in the filaggrin (area of labeling)/nuclei (area of labeling) ratio is evidence of an increase in the differentiation of the epidermal keratinocytes, and consequently of reinforcement of the barrier effect of the epidermis, which constitutes one of the factors of the reinforcement of the state of moisturization of the skin. An increase in keratinocyte differentiation makes it possible to reinforce the barrier effect of the human epidermis, and consequently to decrease the unesthetic effects that may be caused by the dehydration of human skin.

TABLE-US-00010 TABLE 9 Filaggrin (area of labeling)/ nuclei (area of labeling) % stimulation vs control Control medium (culture medium in the absence oftreatment composition) 0.0034 ± 0.0003 — Medium treated with positive reference CaCl.sub.2 1 mMol/L 0.0245 ± 0.0045  621* Medium treat with Composition (C.sub.1A) 0.08% by mass 0.0254 ± 0.0030 648 Medium treated with Composition (C.sub.1Comp) 0.08% by mass Filaggrin (area of labeling)/ % stimulation vs Composition nuclei (area of labeling) (C.sub.1A) 0.0144 ± 0.0059 323p = 0.06

B.2.3. Conclusions

[0191] The application of Composition (C.sub.1A) according to the invention makes it possible to achieve a filaggrin/nucleus ratio value of 0.0254, as opposed to 0.0144 when Composition (C.sub.1A) is replaced with Composition (C.sub.1comp). Thus, the application of Composition (C.sub.1A) according to the invention makes it possible to increase the filaggrin production by 648% relative to the control (untreated keratinocytes) whereas comparative composition (C.sub.1Comp) increases the filaggrin production by only 323%.

[0192] Composition (C.sub.1A) according to the invention makes it possible to increase the keratinocyte differentiation, and consequently to reinforce the barrier effect of the epidermis, and consequently to promote the state of moisturization of said human epidermis.

B.sub.3) Effect of Composition (C.sub.1A) According to the Invention on the Degree of Moisturization of the Skin.

B.3.1. Principle of the Method

[0193] The degree of skin moisturization is determined by evaluating the electrical properties of the skin, for instance the impedance, the resistance and the capacitance, since these dielectric parameters measured at the surface of the skin vary with the amount of water contained in the stratum corneum.

[0194] The principle adopted and used in the context of the present patent application is based on measurement of the variation in the dielectric capacitance of the surface of the skin. Specifically, just like any material or any biological matrix, the stratum corneum can be characterized by its mean capacitance value; this dielectric property varies with the amount of water it contains.

B.3.2 Equipment

[0195] The degree of skin moisturization is measured using a CM825™ model corneometer, sold by the company Courage & Khasaka, equipped with a sensor composed of two metal electrodes. When the corneometer is supplied with electricity, it enables an electric field to be applied across the stratum corneum and measurement of the capacitance corresponding to the state of the skin onto which the electric field has been applied.

B.3.3 Experimental Protocol

[0196] Study of the moisturizing effect of the test compositions was performed on a group of 21 volunteers, and consisted in applying twice a day the compositions to be evaluated, either the placebo (referred to herein as PLAC) or the test composition (referred to herein as COMP), or by leaving an untreated zone (referred to herein as NT), for 21 days. The measurements of the degree of skin moisturization measured using the corneometer were taken before applying the compositions (D0), after a period of seven days following the application of the test compositions to the skin (D7) and after a period of 21 days following the application of the test compositions to the skin (D21).

[0197] The characteristics of the group of volunteers were as follows: [0198] women from 18 to 48 years old, [0199] with an average age of 35, [0200] and of phototype II to IV [0201] and having very dry skin on the legs (≤30 (au) by corneometry on inclusion into the study).

B.3.4 Expressing the Results

[0202] The measurements of the degree of moisturization are expressed in arbitrary units (au).

[0203] An increase in the values (expressed in au) indicates an increase in the water content of the stratum corneum, thus characterizing a moisturizing effect.

[0204] The values measured with the corneometer at each of the measuring times are recorded, and the variations between DO and D7 and then between DO and D21 are expressed as a percentage relative to the value measured at DO, for each composition to be evaluated.

The following are thus defined:
T.sub.(D0): The mean value of the degree of moisturization, expressed in arbitrary units (au), measured before applying the test compositions, on the area to be treated, and on the untreated area;
T.sub.(D7): The mean value of the degree of moisturization, expressed in arbitrary units (au), measured seven days after applying the test compositions, on the treated area, and on the untreated area;
T.sub.(D21): The mean value of the degree of moisturization, expressed in arbitrary units (au), measured 21 days after applying the test compositions, on the treated area, and on the untreated area;

[0205] Δ.sub.7, the percentage increase in the degree of moisturization after seven days of treatment, is thus calculated:

For the placebo:

Δ.sub.7=100×[(T.sub.(D7)−T.sub.(D0)).sub.PLAC−(T.sub.(D7)−T.sub.(D0)).sub.NT]/[T.sub.(D0)).sub.PLAC−(T.sub.(D7)−T.sub.(D0)).sub.NT]

For the test composition:

Δ.sub.7=100×[(T.sub.(D7)−T.sub.(D0)).sub.COMP−(T.sub.(D7)−T.sub.(D0)).sub.NT]/[T.sub.(D0)).sub.COMP−(T.sub.(D7)−T.sub.(D0)).sub.NT]

Δ.sub.21, the percentage increase in the degree of moisturization after 21 days of treatment, is also calculated:
For the placebo:

Δ.sub.21=100×[(T.sub.(D21)−T.sub.(D0)).sub.PLAC−(T.sub.(D21)−T.sub.(D0)).sub.NT]/[T.sub.(D0)).sub.PLAC−(T.sub.(D21)−T.sub.(D0)).sub.NT]

For the test composition:

Δ.sub.21=100×[(T.sub.(D21)−T.sub.(D0)).sub.COMP−(T.sub.(D21)−T.sub.(D0)).sub.NT]/[T.sub.(D0)).sub.COMP−(T.sub.(D21)−T.sub.(D0)).sub.NT]

The effect between the products is also calculated:
% vs placebo at D.sub.7=

100×(T.sub.(D7)−T.sub.(D0)).sub.COMP−(T.sub.(D7)−T.sub.(D0)).sub.PLAC]/[T.sub.(D7)−T.sub.(D0)).sub.PLAC−T.sub.(D7)−T.sub.(D0)).sub.NT]

% vs placebo at D.sub.21=

100×[(T.sub.(D21)−T.sub.(D0)).sub.COMP−(T.sub.(D21)−T.sub.(D0)).sub.PLAC]/[T.sub.(D21)−T.sub.(D0)).sub.PLAC−T.sub.(D21)−T.sub.(D0)).sub.NT]

B.3.5 Results Obtained

[0206] The mean measurements of the degree of moisturization that were obtained for the application of the test compositions are indicated in table 10 below:

TABLE-US-00011 TABLE 10 Product Δ.sub.7 Δ.sub.21 Placebo +18.3% +22.2% Formula containing Composition (C.sub.1A) at 1% by mass +28.8% +37.9%

[0207] The comparisons of the degrees of moisturization obtained between the placebo and the test composition at D7 and D21 are indicated in table 11 below:

TABLE-US-00012 TABLE 11 % vs placebo Product at D 7 at D 21 Formula containing Composition (C.sub.1A) at 1% by mass +58% +56%

B.3.6 Analysis of the Results

[0208] After 7 days following the application of the test compositions, the evolution of the mean of the measured degrees of moisturization Δ7, shows that the increase in the degree of moisturization is 28.8% for composition (C.sub.1A) according to the invention as opposed to 18.8% for the placebo composition. The increase in the degree of moisturization for composition (C.sub.1A) according to the invention is 58% versus placebo.

[0209] After 21 days following the application of the test compositions, the evolution of the mean of the measured degrees of moisturization Δ.sub.21, shows that the increase in the degree of moisturization is 37.9% for composition (C.sub.1A) according to the invention as opposed to 22.2% for the placebo composition. The increase in the degree of moisturization for composition (C.sub.1A) according to the invention is 56% versus placebo.

[0210] As a result, composition (C.sub.1A) according to the invention makes it possible to improve the degree of moisturization of human epidermis.

B.4 Analysis

[0211] The set of results obtained in section B.3 of the present patent application demonstrates that Composition (C.sub.1A) according to the invention affords a moisturizing effect on the epidermis of human skin, both via its role in reinforcing the barrier effect of the epidermis (protection against degradation, and increase) and via its action on increasing the degree of moisturization of the epidermis of human skin.

C) Formulations

[0212] In the following formulas, the percentages are expressed by weight of the formulation.

C.1) -Makeup-Removing Fluid for the Face

Formula

[0213]

TABLE-US-00013 Composition (C.sub.1A) 10.00% Methyl paraben 0.15% Phenoxyethanol 0.80% Sepicalm ™ S 1.00% Perfume/Fragrance 0.10% Water qs 100.00%
Procedure: Mix the various ingredients in the water with magnetic stirring, in the order indicated, and adjust the pH to about 7.

C.2) -Infant Hair and Body Shampoo

Formula

[0214]

TABLE-US-00014 A Composition (C.sub.1A) 15.00% Proteol ™ APL 5.00% Sepicide ™ HB 0.50% Perfume/Fragrance 0.10% B Water 20.00% Capigel ™ 98 3.50% C Water qs 100.00% Sepicide ™ CI 0.30% Colorant qs Sodium hydroxide qs pH = 7.2
Procedure: Mix composition (E.sub.4) with the Proteol™ APL and the Sepicide™ HB (Phase A). Dilute the Capigel™ 98 in a portion of the water and add it to phase A obtained previously (Phase B). Add the rest of the water to phase B, followed by the Sepicide™ CI and the colorant. Adjust the pH of the mixture to about 7.2 with sodium hydroxide.

C.3) -Makeup-Removing Wipes for the Eyes

Formula

[0215]

TABLE-US-00015 A Composition (C.sub.1A) 3.00% B Sepicide ™ HB2 0.50% C Sepicalm ™ VG 0.50% Perfume/Fragrance 0.05% D Water qs 100.00%
Procedure: Mix the ingredients of phase B and those of phase C in phase A until the solution is clear. Add phase D.

C.4) -Mild Foaming Gel

Formula

[0216]

TABLE-US-00016 A Composition (C.sub.1A) 8.50% Proteol ™ APL 3.00% Euxyl ™ PE9010 1.00% Perfume/Fragrance 0.10% B Water qs 100.00% Lactic acid qs pH = 6.0

[0217] Procedure: Dissolve the perfume and the preserving agent Euxyl™ PE9010 in the mixture composed of composition E.sub.4 and the Proteol™ APL (phase A). Add the water and adjust the pH to about 6.0 with lactic acid.

C.5) -Regular-Use Shampoo

Formula

[0218]

TABLE-US-00017 A Composition (C.sub.1A) 12.80% Proteol ™ OAT 5.00% Euxyl ™ PE9010 1.00% Perfume/Fragrance 0.30% Water qs 100.00% B Montaline ™ C40 8.50% Lactic acid qs pH = 6.0
Procedure: Mix all the ingredients of phase A and, after homogenization, add the Montaline™ C40 and adjust the pH to about 6.0 with lactic acid.

C.6 Ultra-Mild Baby Shampoo

Formula

[0219]

TABLE-US-00018 A Composition (C.sub.1A) 10.00% Amisoft ™ CS-11 4.00% Perfume/Fragrance 0.10% Sepicide ™ HB 0.30% Sepicide ™ CI 0.20% Water qs 100.00% B Water 20.00% Capigel ™ 98 3.50% Tromethamine qs pH = 7.2
Procedure: Mix all the ingredients of phase A, in the order indicated, until a clear phase A is obtained. Separately, add the Capigel™ 98 to the water and then add this phase B thus prepared to phase A and adjust the pH to 7.2 using tromethamine.

C.7 Baby Cleansing Milk

Formula

[0220]

TABLE-US-00019 A Simulsol ™ 165 2.00% Montanov ™ 202 1.00% Lanol ™ 99 3.00% Dimethicone 1.00% Isohexadecane 3.00% B Water qs 100.00% C Sepiplus ™ 400 0.30% D Composition (C.sub.1A) 6.35% E Sepicide ™ HB 0.30% DMDM Hydantoin 0.20% Perfume/Fragrance 0.10%
Procedure: Heat, separately, phases A and B constituted by mixing the various constituents. Add phase C to the hot fatty phase and make the emulsion by pouring in the aqueous phase; homogenize for a few minutes with vigorous stirring (by means of a rotor/stator turbomixer). Next, add phase D to the hot emulsion and cool the emulsion with moderate stirring down to room temperature. Add phase E at 40° C.

C.8 Cleansing Powder Lotion for Sensitive Skin

Formula

[0221]

TABLE-US-00020 A Lipacide ™ C8G 0.95% Methyl paraben 0.10% Ethyl paraben 0.024% Propyl paraben 0.0119% Butyl paraben 0.024% Isobutyl paraben 0.0119% Water 20.00% Disodium EDTA 0.10% Triethanolamine 1.38% B Composition (C.sub.1A) 1.80% Perfume/Fragrance 0.10% C Sepicalm ™ S 0.28% Water qs 100.00% Lactic acid qs pH = 5.2 D Micropearl ™ M310 5.00%

[0222] Procedure: Dissolve the ingredients of phase A in the water at 80° C. Separately, dissolve the fragrance in composition (E.sub.4) to prepare phase B. Add the cooled phase A to phase B, then introduce the Sepicalm™ S and the remaining water. Check the final pH and adjust to about 5.2 if necessary. Next, add the Micropearl™ M310.

C.9 Infant Shower Gel

Formula

[0223]

TABLE-US-00021 A Water 56.06% Sepimax ™ Zen 3.00% Sepiplus ™ S 0.80% B Proteol ™ OAT 20.80% Oramix ™ NS 10 9.30% Amonyl ™ 265 BA 5.10% C Composition (C.sub.1A) 2.00% Glyceryl glucoside 1.00% Phenoxyethanol & ethylhexyl glycerol 1.00% Perfume/Fragrance 0.90% Colorant 0.04%
Procedure: disperse the Sepimax™ ZEN in the water and stir using a mechanical stirrer equipped with a deflocculator, a counter-rotating impeller and an anchor paddle, until a perfectly smooth gel is obtained. Add the Sepiplus™ S and then stir until the mixture is homogeneous. Next, add the ingredients of phase B, homogenize and individually add the additives of phase C. Adjust the pH to 6.0-6.5.

C.10BB Cream

Formula

[0224]

TABLE-US-00022 A Easynov ™ 2.30% Lanol ™ 99 1.00% Sepimat ™ H10W 1.00% Ethylhexyl methoxycinnamate 5.00% B Cyclomethicone 6.00% Triethoxycaprylsilane & alumina-silane & titanium oxide 8.00% Red iron oxide & triethoxycaprylsilane 0.24% Yellow iron oxide & triethoxycaprylsilane 0.66% Black iron oxide & triethoxycaprylsilane 0.09% Perfume/Fragrance 0.10% C Water qs  100% Sepinov ™ EMT10 1.20% D Composition (C.sub.1A) 2.00% Sepitonic ™ M3 1.00% Phenoxyethanol & ethylhexyl glycerol 1.00%
Procedure: Prepare phase B by mixing the various ingredients and homogenize using a mixer equipped with a rotor-stator system at a spin speed of 4500 rpm, for a period of 6 minutes. Prepare phase C by adding the Sepinov™ EMT10 to the mixture of water and glycerol, and homogenize using a mixer equipped with a rotor-stator system at a spin speed of 4000 rpm for 4 minutes. Add phases A and B to phase C, and stir the resulting mixture using a mechanical stirrer equipped with an anchor paddle, at a speed of 30 rpm for 2 minutes, and then at a speed of 50 rpm for 20 minutes. Add the components of phase 5 one by one and stir at a speed of 50 rpm for 25 minutes.
C.11 High-Protection Antisun Spray with an SPF of Greater than 30

Formula

[0225]

TABLE-US-00023 A Montanov ™L 1.00% Montanov ™ 82 1.00% C12-15 Alkyl benzoate 17.00% Dimethicone 3.00% Octocrylene 6.00% Ethylhexyl methoxycinnamate 6.00% Bis(ethylhexyloxyphenol)methoxyphenyltriazine 3.00% Tocopherol 0.05% B Water qs 100% C Simulgel ™ INS 100 0.50% Cyclodimethicone 5.00% D Composition (C.sub.1A) 3.00% Phenoxyethanol & ethylhexyl glycerol 1.00% Perfume/Fragrance 0.20% E Methylenebis(benzotriazolyl) Tetramethylbutylphenol 10.00% 25% Citric acid qs pH = 5
Sepicalm™ S: Mixture of N-cocoyl amino acids, sarcosine, potassium aspartate and magnesium aspartate as described in WO 98/09611;
Proteol™ APL: Mixture of sodium salts of N-cocoyl amino acids, obtained by acylation of amino acids characteristic of apple juice;
Sepicide™ HB: Mixture of phenoxyethanol, methyl paraben, ethyl paraben, propyl paraben and butyl paraben, which is a preserving agent;
Capigel™ 98: Acrylate copolymer;
Sepicide™ CI: Imidazoline urea, which is a preserving agent;
Sepicide™ HB: Mixture of phenoxyethanol, methyl paraben, ethyl paraben, propyl paraben, butyl paraben and isobutyl paraben, which is a preserving agent;
Sepicalm™ VG: Mixture of N-palmitoyl proline in sodium salt form and of extract of Nymphea alba blossom;
Euxyl™ PE9010: Mixture of phenoxyethanol and ethylhexyl glycerol;
Proteol™ OAT: Mixture of N-lauryl amino acids obtained by total hydrolysis of oat protein as described in WO 94/26694;
Montaline™ C40: Chloride salt of monoethanolamine cocamidopropyl betainamide;
Amisoft™ CS-11: Sodium salt of N-cocoyl glutamate;
Simulsol™ 165: Mixture of PEG-100 stearate and glyceryl stearate;
Montanov™ 202 (arachidyl alcohol, behenyl alcohol and arachidyl glucoside) is a self-emulsifying composition such as those described in EP 0 977 626;
Lanol™99: Isononyl isononanoate;
Sepiplus™ 400: Self-invertible inverse latex of polyacrylates in polyisobutene and including polysorbate 20, as described in WO 2005/040230;
Lipacide™ C8G: Capryloylglycine sold by the company SEPPIC;
Micropearl™ M310: Crosslinked polymethyl methacrylate polymer in powder form, used as a texture modifier;
Sepimax™ Zen (INCI name: Polyacrylate Crosspolymer-6): Thickening polymer in the form of a powder;
Sepiplus™ S (INCI name: Hydroxyethyl Acrylate/Sodium Acryloyldimethyl Taurate Copolymer & Polyisobutene & PEG-7 Trimethylolpropane Coconut Ether): Self-invertible inverse latex;
Amonyl™ 265 BA (INCI name: cocoyl betaine): Foaming amphoteric surfactant;
Sepinov™ EMT10 (INCI name: Hydroxyethyl Acrylate/Sodium Acryloyldimethyl Taurate Copolymer): Thickening copolymer in the form of a powder;
Easynov™ (INCI name: Octyldodecanol and Octyldodecyl Xyloside and PEG-30 Dipolyhydroxystearate): Emulsifying agent of lipophilic tendency;
Sepimat™ H10 FW (INCI name: Methyl Methacrylate Crosspolymer and Squalane): Polymer used as texture agent;
Sepitonic™ M3 (INCI name: Magnesium Aspartate and Zinc Gluconate and Copper Gluconate): Mixture used as free-radical scavenger and energizing agent for cells;
Montanov™ L (INCI name: C14-22 Alcohols and C12-20 Alkylglucoside): Emulsifying agent;
Montanov™ 82 (INCI name: Cetearyl Alcohol and Coco-glucoside): Emulsifying agent;
Simulgel™ INS100 (INCI name: Hydroxyethyl Acrylate/Sodium Acryloyldimethyl Taurate Copolymer and isohexadecane and Polysorbate 60): Polymeric thickener.