WO2011110793A2

WO2011110793A2 - Cosmetic firming active principle from piper nigrum, cosmetic compositions containing same and use thereof

Info

Publication number: WO2011110793A2
Application number: PCT/FR2011/050498
Authority: WO
Inventors: Jean Paufique
Original assignee: Societe Industrielle Limousine D'application Biologique, Dite Silab
Priority date: 2010-03-12
Filing date: 2011-03-11
Publication date: 2011-09-15
Also published as: FR2957259A1; WO2011110793A3; FR2957259B1

Abstract

The invention relates to the use of an active principle originating from Piper nigrum as a cosmetic firming active principle, particularly to combat laxity in photo-aged skin. The invention also relates to a specific active principle originating from Piper nigrum berries, as well as to cosmetic compositions containing said active principle and to a cosmetic skin-care method for firming the skin, in particular acting on the functionalities and the synthesis capabilities of the reticular dermis.

Description

ACTIVE REFINING COSMETIC PRINCIPLE FROM PIPER NIGRUM, COSMETIC COMPOSITIONS INCLUDING AND

USE

The present invention relates to the cosmetic use of a Piper nigrum extract as a firming active ingredient, as well as to a cosmetic skin care process for firming the skin, in particular for combating sagging skin and for strengthening the tone and firmness of aged photo skin.

The invention also relates to a particular cosmetic active ingredient derived from Piper nigrum and to cosmetic compositions containing this active ingredient.

The aging process of the skin is complex and results from both intrinsic aging or chronological aging and extrinsic aging or environmental aging.

However, environmental aging represents the major part of skin aging and is mainly due to UV (ultraviolet) irradiation which leads to alterations of the proteins of the dermal matrix, thus causing the formation of wrinkles and the loss of the elasticity properties of the tissue.

The dermis is subdivided into two zones.

The first, superficial and fine, is located between the dermal papillae and called papillary dermis. It is formed of loose connective tissue with a high density of fibroblasts.

The second, deeper, is the reticular dermis. Thicker, it is formed of a dense matrix of compact collagen bundles crisscrossed and intermixed with elastic fibers growing thicker towards the depth of the dermis. These Fibrous molecules are predominant on fibroblasts and the amorphous ground substance.

The reticular dermis and the papillary dermis possess fibroblasts with distinct morphological characteristics and biological properties. The reticular dermis and the papillary dermis also differ in the composition of their respective extracellular matrix. The reticular fibroblasts produce more matrix components than the papillary fibroblasts. Studies have also highlighted differences in expression, synthesis or deposition of structural and matricellular proteins, as well as growth factors and degradation enzymes.

Chronobiological aging is known to have little effect on the reticular dermis, while on the other hand it is sensitive to extrinsic aging. After UV irradiation, the functionalities of the reticular dermis and its capacities of synthesis are altered: the dermis then loses of its substance and its structure leading to a strong cutaneous relaxation.

Therefore, the objective of the present invention is to provide an effective firming cosmetic solution capable of fighting against the sagging of the cutaneous tissue and against the relaxation of the face to redraw and rejuvenate the features, in particular capable of preserving or restoring the functionalities and synthetic capacities of the reticular dermis.

For this purpose the invention proposes to use an asset from Piper nigrum.

Piper nigrum or black pepper is one of 700 species of Piper. It belongs to the family Piperaceae. Its fruits also called berries are mainly used in food as a spice.

Surprisingly, Piper nigrum has noticeable effects on the cutaneous level with very interesting cosmetic properties for firming the skin, in particular action capacities at the level of the reticular dermis. The present invention therefore aims to use at least one extract derived from Piper nigrum as a firming cosmetic active ingredient.

By "extract derived from Piper nigrum" or "active ingredient from Piper nigrum" within the meaning of the present invention means any aqueous extract, glycolic or hydrolyzate obtained from Piper nigrum.

Preferably, the invention aims to use an extract derived from Piper nigrum berries.

Likewise, in a preferred manner, the invention aims to use an oligosaccharide extract derived from Piper nigrum.

For the purposes of the present invention, the term "oligosaccharide extract" or "oligosaccharide active principle" means an extract or active principle comprising oligosaccharides. It is preferably an extract comprising at least 20% of saccharides of which at least 50% in the form of oligosaccharides. Preferably, the oligosaccharides present are predominantly oligosaccharides with a degree of polymerization of between 2 and 7.

The invention also relates to a cosmetic active ingredient derived from Piper nigrum berries comprising in particular oligosaccharides with a degree of polymerization of between 2 and 7, as well as cosmetic compositions including at least one such active principle.

The active ingredients according to the invention are preferably hydrolysates. The cosmetic compositions according to the invention contain from 0.01 to 20% of an active ingredient derived from Piper nigrum berries comprising oligosaccharides.

The administration of a composition containing an active ingredient derived from Piper nigrum according to the invention is carried out topically. The compositions according to the invention may be in any form allowing the application topically. Finally, the invention also relates to a cosmetic skin care method, intended to firm the skin, comprising the topical application of a composition containing an active ingredient from Piper nigrum.

The present invention is now described in detail.

USE

The invention thus relates to the use of at least one extract derived from Piper nigrum as a cutaneous firming cosmetic active ingredient.

According to the invention, an active ingredient derived from Piper nigrum can indeed be used as a cosmetic agent on the skin to firm the cutaneous tissues.

The presence of oligosaccharides with a degree of polymerization of between 2 and 7 is preferred and makes it possible in particular to confer on it this cosmetic activity. Preferably the invention is directed to the use of an oligosaccharide extract derived from Piper nigrum, comprising α-glucans and rhamnogalacturonans. Advantageously, the use on the skin of an active ingredient derived from Piper nigrum promotes the fight against sagging skin tissues and / or to increase the tone and firmness of the skin and / or to fight against slackening. facial skin. It allows to redraw and rejuvenate facial features.

The active ingredient according to the invention can specifically be used in a cosmetic composition, to act on the reticular dermis. The active ingredient and / or the composition may in particular be intended to act on the functionalities and synthetic capacities of the reticular dermis, in particular to preserve these functionalities and synthesis capabilities in the face of UVA irradiation.

Indeed, according to a first aspect of the invention, an extract of Piper nigrum may be used to enhance the functionality of the reticular dermis, in particular:

- its membership capabilities, - his contraction abilities, and

- its migration capabilities.

Membership capabilities

Cell adhesion corresponds to the set of cellular and molecular mechanisms used to adhere the cells together or with their own microenvironment via dynamic interactions that are both stable and variable. The specific interactions developed between the fibroblasts and the collagen network, mediated by the α2β1 integrins, give rise to a functional tissue ensuring the maintenance of the upper layers of the skin, its strength and resistance, and also conditions the reciprocal exchanges between cells and the extracellular matrix.

Adhesion abilities can be particularly affected during extrinsic skin aging. This reduction in the adhesion capacities of the reticular fibroblasts, results in a loosening of the skin.

According to the invention, an active ingredient derived from Piper nigrum may be used to improve or preserve the adhesion capacities of the reticular fibroblasts, in particular to restore the adhesion capacities of photo-aged reticular fibroblasts.

Contractility

The reticular fibroblasts, thanks to their high adhesion capacity, anchor to the extracellular matrix and develop contractile properties via the reorganization of the filaments of their cytoskeleton. These contractile properties generate tensile forces that stimulate the organization and retraction of surrounding collagen fibers essential for continuous remodeling of the extracellular matrix. This process is necessary for the generation of skin's resistance properties.

Studies of the Applicant have shown that the contractile properties of the reticular fibroblasts can be affected in particular by aging. extrinsic of the skin. In particular, these collagen lattice studies have shown that the contraction capacities of the photoagedillion fibroblasts were significantly reduced compared with non-photo-aged reticular fibroblasts. Indeed, the photo-aging is accompanied by fragmented collagen fibers, and less fibroblast adhesion capacity does not preserve the extracellular matrix under tension.

This alteration of the contractile properties of the reticular fibroblasts results in a relaxed skin, which loses tone and firmness.

According to the invention, an active ingredient derived from Piper nigrum can be used to improve or preserve the contraction capacities of reticular fibroblasts, in particular to restore the contractile power of photo-aged reticular fibroblasts.

Migration capabilities

Cell migration is an essential process for various biological events, particularly for the regeneration of damaged tissues. It is a dynamic process that includes the polarization and progressive movement of fibroblasts in a given orientation. The migration of fibroblasts is strongly modulated by the chemical characteristics (growth factors and chemo-attractant factors), and by the physical characteristics (density, rigidity, tension) of the micro matrix environment.

The migration capacity of the fibroblasts of the reticular dermis is essential to regenerate the damaged dermis. In fact, excessive skin damage, particularly UVA irradiation, can lead to the disappearance of papillary fibroblasts by apoptosis, and the reticular fibroblasts that survive, thanks to their plasticity and their high dynamics, are able to progressively migrate towards the skin. upper part of the dermis to replace them. According to the invention, an active ingredient derived from Piper nigrum can be used to stimulate the migration capacities of the reticular fibroblasts, so as to be able to regenerate the dermas assaulted. According to another aspect of the invention, a Piper nigrum extract may also be used to enhance the expression capacities of the matrix proteins of the reticular fibroblasts, in particular:

to stimulate the expression of fibrillar molecules, in particular collagen I and elastin,

to stimulate the expression of the adhesion molecules, in particular the α2β1 integrin, and

to stimulate the expression of the signaling matrix molecules, in particular the CT & F (Connective tissue growth factor) growth factor. Fibrillar molecules

The reticular dermis is woven in its full depth with a large three-dimensional network of intertwined thick fibers of intertwined collagen and elastin that give the tissue its architecture and biomechanical properties.

The co-distribution and close assembly of different types of collagen form a complex scaffold that stabilizes the structure of the skin and ensures the biomechanical properties of the tissue. In particular, collagens I, III and V or fibrillar collagens are arranged in a particular way to define the specific architecture of the dermis and to confer on the skin its biomechanical properties of density and resistance.

Moreover, the reticular dermis is characterized by elastic fibers, formed of the assembly of tropoelastine monomers and stabilized under the effect of lysyl oxidase, which, because of their particular structure, can be strongly stretched and ensure the skin its elasticity. When syntheses of collagen, especially collagen I, and elastin are affected, this results in an alteration of the biomechanical properties of the skin. These syntheses are notably diminished during extrinsic skin aging. Extrinsic aging particularly affects the genes encoding the structural and remodeling proteins of the extracellular matrix. UV irradiations induce the reduction of collagen I syntheses as well as the fragmentation of fibers leading to collagen depletion and alteration of the biomechanical properties of the skin. In addition, photoaging is characterized by a massive accumulation of truncated and nonfunctional elastic fibers forming amorphous masses in the reticular dermis. The skin tissue loses its structure and density and relaxes.

According to the invention, an active ingredient derived from Piper nigrum can be used to stimulate collagen I and / or elastin synthesis, in particular to restore the expression of collagen I and / or elastin in photochromic reticular fibroblasts. old.

Adhesion molecules: integers

Integrins are transmembrane receptors specialized in cell-cell and extracellular matrix-cell interactions. They are able to transmit signals bidirectionally to the plasma membrane, thus controlling cell behavior and tissue organization.

Among the integrins, the α2β1 integrin is specifically involved in the attachment of fibroblasts to collagen fibers. It modulates many signaling pathways and regulates various cellular processes including proliferation, survival and migration. It also plays a role in remodeling collagen fibers and contraction of tissue. A decrease in α2β1 integrins leads to the loss of their structural and signaling role necessary for the cohesion of the cutaneous tissue and the mutual exchanges fibroblasts - extracellular matrix, and the skin loses its tonicity and its firmness. This is particularly the case when the skin is attacked, especially during UV irradiations.

According to the invention, an active ingredient derived from Piper nigrum can be used to stimulate the synthesis of α2β1 integrin, in particular to restore the expression of this molecule in photo-aged reticular fibroblasts, and thus to fight against sagging skin.

Matrix signaling molecules: CT & F or CCN2

CT & F or CCN2 is a matricellular protein for interaction and adhesion between fibroblasts and the extracellular matrix, which plays an important role in signal transduction. The reticular fibroblasts express more CT & F than the papillary dermis cells.

CT & F participates in the regulation of matrix syntheses by intervening in the signaling pathways of growth factors. In particular, he mediates the inducing effects of TGF-pi on the synthesis of collagen I.

It also promotes the adhesion of fibroblasts to its matrix as a molecular bridge between matrix proteins and their receptors on the surface of fibroblasts. In addition, by binding to fibroblasts through integrins or proteoglycans, it controls the reorganization of the cytoskeleton and the formation of lamellipodia and thus regulates the ability of fibroblasts to migrate or expand in the matrix.

A reduction in the expression of CT & F, for example induced in the case of extrinsic aging, contributes to the reduction of procollagen synthesis and affects the interaction, migration and adhesion capabilities of the dermis: the skin tissue collapses and the skin relaxes. According to the invention an active ingredient derived from Piper nigrum can be used to stimulate the synthesis of CT & F, in particular to restore the expression of CT & F in the photo-aged reticular fibroblasts, and thus firm the skin.

An extract derived from Piper nigrum can therefore be used as a cosmetic active ingredient in a cosmetic composition to preserve the adhesion, contraction and migration capacities of fibroblasts and their synthesis capacities, primary functions of the reticular dermis and targets. photo-aging, thus reinforcing the structure, strength and support properties of the skin for a firming effect. It can in particular be used as a cosmetic active ingredient inducing the synthesis of collagen I, elastin, α2β1 integrin and / or CT & F in skin cells, and specifically in the reticular dermis.

Advantageously, according to the invention, an extract from Piper nigrum can bring tonicity and firmness to the skin and limit the ptosis of the face, especially for the exposed skin. It corrects the impact of photo-aging on the parameters related to sagging skin and allows people with photo-exposed skin to find the characteristics of a non-photo-exposed skin.

Preferentially, the Piper nigrum extract that is useful according to the invention is the active ingredient described below.

ACTIVE INGREDIENT

The invention also relates to a particular cosmetic active ingredient obtained from Piper Nigrum berries comprising oligosaccharides with a degree of polymerization of between 2 and 7.

Preferably, it is an extract having an oligosaccharide content with a degree of polymerization of between 2 and 7 of at least 10% by total weight of the dry matter, in particular a content of between 10% and 32%. in total weight of the dry matter. Preferably, the oligosaccharides are in the form of α-glucans and rhamnogalacturonans.

The active ingredient according to the invention may also comprise other molecules, in particular proteins, phenolic compounds, uronic acids and minerals.

Preferably, the active ingredient according to the invention does not contain piperine. According to a particularly suitable embodiment, the active ingredient according to the invention is a hydrolyzate of Piper nigrum berries.

The active ingredient according to the invention is preferably in the form of a clear amber liquid. It can be defined by at least one of the features described below, preferably all.

Dry matter:

The dry matter content of an active ingredient according to the invention (measured by passing in an oven at 105 ° C. in the presence of sand of a sample of initial weight given until a constant weight is obtained) can be between

8 and 100 g / l, preferably between 17 and 25 g / l.

PH measurement:

The pH (measured by the potentiometric method at room temperature) can be between 3.0 and 6.0, preferably between 3.0 and 4.0. Carbohydrates:

Determination of total sugar content:

The determination of the total sugar content can be carried out by the DUBOIS method (DUBOIS M. et al., (1956), Analytical Chemistry, 28, No. 3, pp. 350-356). In the presence of concentrated sulfuric acid and phenol, the reducing sugars give an orange-yellow compound. From a standard range, the total sugar content of a sample can be determined. The total sugar level of an active ingredient from Piper nigrum according to the invention is preferably between 2 and 36 g / l, more preferably between 5 and 9 g / l.

The total sugar content can also be expressed as a percentage relative to the dry matter. It is included in 20% and 53%.

Characterization of simple sugars

The characterization of simple sugars is carried out by ionic liquid chromatography.

The level of each sugar is determined by means of a standard range of the synthetic sugar studied.

Thus, preferably, the active ingredient from Piper nigrum according to the invention is composed of the following simple sugars, in descending order: glucose, galactose, galacturonic acid, arabinose, rhamnose, mannose and fructose.

For example, the particular nonlimiting active ingredient used for the illustrative tests of the invention, has the following composition of simple sugars:

Characterization of carbohydrate size

The determination of the carbohydrate size of an active ingredient derived from

Piper nigrum according to the invention is carried out by high performance liquid chromatography. The molar masses of the carbohydrates present in the active principle are presented in the following table.

The carbohydrates of the active principle according to the invention are therefore preferentially distributed as follows: between 40 and 50% of monosaccharides with a molecular mass of less than 180 Da and between 50 and 60% of oligosaccharides and polysaccharides with a molar mass of between 180 and 5220 Da.

The carbohydrate fraction of the active ingredient derived from Piper nigrum according to the invention is therefore essentially composed of glucose present in the form of monosaccharides and oligosaccharides.

By way of example, the molar masses of the carbohydrates present in the particular nonlimiting active principle used for the tests are presented in the following table.

Molar mass (Da)

Rate and degree of polymerization

corresponding carbohydrates (%)

Monosaccharides MM <180 47.7%

Oligosaccharides and

180 <MM <5220 53.3% polysaccharides Characterization of oliqosaccharides

The composition of the oligosaccharides of the active ingredient derived from Piper nigrum was determined by analysis of the products obtained after activity of various enzymes on the active principle.

The majority of oligosaccharides have a degree of polymerization of between 2 and 7.

The oligosaccharides are preferably in the form of α-glucans and rhamnogalacturonans of type I.

The carbohydrate fraction of the active ingredient of Piper nigrum therefore comprises:

oligosaccharides of α-glucans and rhamnogalacturonans of type I, with a degree of polymerization essentially between 2 and 7, and

glucose monosaccharides.

The oligosaccharides preferably represent between 50 and 60% of the total sugars, and the simple sugars between 40 and 50%.

Phenolic compounds:

The content of total polyphenols is measured according to the following principle. The polyphenols form colored compounds in the presence of potassium ferricyanide, the intensity of this coloration being proportional to the amount of polyphenols. By referring to a standard range of gallic acid, in this case from 40 to 120 mg / l, it is possible to deduce the amount of polyphenols present in the extract.

The total polyphenol content of the active ingredient is preferably between 0.3 and 0.5% by total weight of dry matter.

Uronic acids:

The content of uronic acids is determined by the following principle: the product of galacturonic acid with sodium tetraborate, gives in the presence of meta-hydroxydiphenyl, a pink color for assay spectrophotometer at 520 nm.

The intensity of staining is proportional to the amount of uronic acids. Referring to a standard range of galacturonic acid, in this case from 10 to 100 mg / l, it is possible to deduce the quantity of uronic acids present in the active principle.

The uronic acid content of the active ingredient is preferably between 3.2 and 5.0% by total weight of dry matter.

Protein content:

The protein content is determined by the following principle: The total nitrogen is determined by the method of KJELDHAL (reference: Official Method of Analysis of the AOC 1975, 12th ed., W Horwitz, ED, New York, p. 15-60).

The protein content is expressed as a percentage relative to the dry matter.

The protein content of the active ingredient is preferably between 22.7 and 34.1% by total weight of dry matter.

Ash / mineral content:

The raw ash content is determined by the weighing of residues from incineration at 550 ° C in an electric muffle furnace (VULCANTM 3.550-NDI). The weight of the residue is calculated by deducting the tare.

The raw ash / mineral content is expressed as a percentage of the dry matter.

The mineral content of the active ingredient is preferably between 28.4 and 42.8% by total weight of dry matter.

Alkaloid content:

The alkaloid content is determined using Dragendroff's reagent (0.1M potassium iodate, 2.9M glacial acetic acid, 1.5mM bismuth subnitrate), in visual comparison with a range of quinine. The presence of a red precipitate shows the presence of alkaloid and the absence of red precipitate shows the absence of alkaloid (amount less than the sensitivity threshold of this method, ie less than 0.05 g / l). Very preferably, with the active principle according to the invention, there is an absence of red precipitate. According to a preferred embodiment, the active principle according to the invention therefore does not contain alkaloid, so no piperine.

Identification of the active fraction

In order to determine the predominantly active fraction of an active ingredient according to the invention, a study has been carried out. This study consists in determining the main fraction of the active principle according to the invention acting on the contractile power of human fibroblasts reticular.

For this study, the molecular species (neutral carbohydrates, peptides and minerals) of the active ingredient from Piper nigrum were fractionated by solid phase extraction:

Fraction A: The neutral carbohydrates are purified by successive adsorption of the cationic and then anionic compounds on ionic resins.

Fraction B: The peptides are purified by adsorption / elution on a cation exchange resin.

Fraction C: The ash fraction is obtained by resolubilization of residues from incineration in an electric muffle furnace.

The results obtained show that only fraction A has an efficacy on the contractile power of human fibroblasts reticular similar to that of the active ingredient from Piper nigrum.

Fraction A consists mainly of sugar, essentially in the form of oligosaccharides with a degree of polymerization predominantly between 2 and 7.

The major active fraction of the cosmetic active ingredient according to the invention therefore consists of oligosaccharides, with a degree of polymerization predominantly between 2 and 7. PROCESS FOR OBTAINING

The active ingredient according to the invention can be obtained preferentially by enzymatic hydrolysis.

A particularly suitable method comprises at least the succession of the following steps:

aqueous solubilization of Piper nigrum berry powder,

enzymatic hydrolysis making it possible to obtain, at least partly, oligosaccharides with a degree of polymerization of between 2 and 7,

separation of the soluble and insoluble phases,

- Enzymatic inactivation by heat treatment,

- f iltrations and / or sterilizing filtration.

Decolorization and deodorization steps of the soluble phase can be envisaged without modifying the active part of the active ingredients.

The various steps are customary in the field of extraction of active ingredients from plants, and the skilled person is able to adjust the reaction parameters on the basis of his general knowledge, in order to obtain extracts containing the less oligosaccharides.

COSMETIC COMPOSITIONS AND COSMETIC PROCESS FOR SKIN CARE

The present invention also covers cosmetic compositions including at least one active ingredient derived from Piper nigrum berries in different galenic forms, adapted for topical administration to the skin.

These compositions may be in the form of oil-in-water emulsions, water-in-oil emulsions, multiple emulsions (water / oil / water or oil / water / oil) which may be microemulsions or nanoemulsions, or in the form of solutions, suspensions, hydrodispersion, aqueous gels or powders. They can be more or less fluid and look cream, lotion, milk, serum, ointment, gel, paste or foam, or in solid form.

These compositions contain between 0.01% and 50%, in particular between 0.01% and 20%, by weight of active principle (s) derived from Piper nigrum berries according to the present invention, preferably between 0.5% by weight. % and 3%.

These compositions comprise, in addition to the active agent, a physiologically acceptable and preferably cosmetically acceptable medium, that is to say which does not cause unacceptable sensations of discomfort for the user such as redness, tightness or tingling. This medium usually contains water.

The compositions according to the invention may contain as adjuvant at least one compound chosen from:

oils, which may be chosen in particular from silicone oils, linear or cyclic, volatile or non-volatile;

waxes, such as ozokerite, polyethylene wax, beeswax or carnauba wax;

silicone elastomers,

surfactants, preferably emulsifiers, whether they are nonionic, anionic, cationic or amphoteric;

co-surfactants, such as linear fatty alcohols;

thickeners and / or gelling agents,

humectants, such as polyols such as glycerine;

- organic filters,

inorganic filters,

- dyes, preservatives, fillers,

- the tensors,

- the sequestering agents,

- the perfumes, - and their mixtures, without this list being limiting.

Examples of such adjuvants are cited in particular in the CTFA Dictionary (International Cosmetic Ingredient Dictionary and Handbook published by the Personal Care Product Council).

These compositions are especially intended for the firming of human skin, the skin of the body or the skin of the face, in particular the care of skins having undergone a photo-aging.

To this end, the invention aims at a cosmetic process for the care of human skin, intended for firming the skin, comprising the topical application of a composition containing an active ingredient derived from Piper nigrum, in particular a composition containing between 0.01 and 20% by weight of active ingredient (s) derived from Piper nigrum.

The invention also relates to a cosmetic process for the care of human skin, intended to combat the loosening of the skin of the face and / or to strengthen the tonicity and firmness of the skin, in particular for the skins which have undergone a photo- aging, comprising the topical application of a composition containing an active ingredient derived from Piper nigrum, in particular a composition containing between 0.01 and 20% by weight of active ingredient (s) derived from Piper nigrum according to the present invention.

Of course, those skilled in the art will take care to choose any additional compounds, active or non-active, and / or their quantity, so that the advantageous properties of the mixture are not, or not substantially, altered by the addition considered. EXAMPLES

A non-limiting example of a process for obtaining an active ingredient obtained from Piper nigrum berries is presented below, as well as examples of compositions including such an active principle. Example 1 Process for Obtaining the Active Ingredient According to the Invention

An example of a process for obtaining an active ingredient according to the invention comprises the implementation of the following steps:

solubilization of 50 g of Piper nigrum berry powder in 1 kg of water, enzymatic hydrolysis by means of a protease,

separation of the soluble and insoluble phases by decantation,

- enzymatic inactivation by heat treatment at 80 ° C for at least 5h,

filtration in order to select oligosaccharides with a degree of polymerization of between 2 and 7,

sterilizing filtration on 0.22μ.

The active ingredient obtained has the following characteristics:

- Clear, amber-colored liquid with a characteristic odor

- Dry matter content: 19.4g / l

- pH = 3.6

- Total sugar content: 6.0g / l, ie 30.9% relative to the dry matter

- Protein content: 5.5g / l, 28.4% relative to the dry matter

- Ash content of 35.6%.

- Total polyphenol content of 0.4%.

- Uronic acid content of 4.1%.

- Absence of alkaloid, therefore absence of piperine.

Example 2 Use of an Active Ingredient of the Invention in an Evanescent Firming Balm

Phase A. Water QSP 100%

Carbopol Ultrez 20 (Noveon) 0.4%

Phase B. Finester EH25 (Cornelius) 7.3%

DUB 340 (Stéarinerie Dubois) 2% DUB PTL (Stéarinerie Dubois) 6%

DUB MM (Stéarinerie Dubois) 3%

DUB PPH1 (Stéarinerie Dubois) 2%

Sophim MC30 (Sophim) 6%

Phase C. Conservative 0.7%

Active principle according to the invention (example 1) 3%

Phase D. NaOH QSP pH 5.5%

The quantities indicated are given as a percentage of weight.

This thick emulsion has a pH of 5.5.

It can be obtained by implementing the following steps:

- Mix A, disperse the gel at 50 ° C with mechanical stirring,

- Mix B, heat A and B at 80 ° C, with mechanical stirring,

Emulsifying B in A under a rotor stator emulsifier,

- At 50 ° C add C,

- At 30 ° C, adjust the pH with D, and leave in an emulsifier until the emulsion is uniform.

Example 3: Use of an active ingredient of the invention in a tonic foam Phase A. Water QSP 100 °

Carbopol ETD2050 (Novéon) 0.4%

Phase B. DUB liquid 85 (Stéarinerie Dubois) 5.7%

Arlatone 2121 (ICI) 3%

Emulgade 1000 NI (Henkel) 1.3% itachol 2000 (Rita) 2.7%

DUB G1218A (Stéarinerie Dubois) 2%

DUB MCT 5545 (Stéarinerie Dubois) 2.3%

Phase C. Conservative 0.7%

Active principle according to the invention (example 1) 3%

Phase D. NaOH QSP pH4.5 The quantities indicated are given as a percentage of weight.

This emulsified foam has a pH of 4.5.

It can be obtained by implementing the following steps:

- Mix A, well disperse the gel at 50 ° C with mechanical stirring, - Mix B, heat A and B at 80 ° C, with mechanical stirring,

Emulsifying B in A under a rotor stator emulsifier,

- At 40 ° C, add C,

At 30 ° C., aerate the emulsion under a stator rotor while adjusting the pH with D, and stirring vigorously until a fine and homogeneous foam is obtained. EXAMPLE 4 Use of an Active Principle of the Invention in a Day Creamy Cream

Phase A. Water QSP 100%

Carbopol Ultrez 20 (Noveon) 1%

Phase B. Lanol 37T (Seppic) 2%

Cutina E24 (Henkel) 3%

DC 7,3101 (Dow Corning) 4%

Pelemol 2014 (Cornelius) 5%

DC 200 (Dow Corning) 1%

DC 345 (Dow Corning) 1%

DUB PTCC (Stéarinerie Dubois) 5%

DUB Synersol (Stéarinerie Dubois) 2%

Phase C. Conservative 0.7%

Active principle according to the invention (example 1) 3%

Phase D. NaOH Qsp pH 4.5 Amounts shown are given as percentages by weight.

This emulsified gel has a pH of 4.5.

It can be obtained by the implementation of the following steps:

- Mix A, disperse the gel well with mechanical stirring, - Mix B,

- Heat A e B at 80 ° C,

Emulsifying B in A under a foam concentrate,

- At 40 ° C add C and homogenize,

- Adjust the pH with D, with strong mechanical stirring, and continue the homogenization in an emulsifier up to 30 ° C.

Example 5 Use of an Active Ingredient of the Invention in a Night Cream

Phase A. Water QSP 100% Phase B. DUB BA (Stéarinerie Dubois) 4%

DUB MCT 5545 (Stéarinerie Dubois) 5%

Stearyl alcohol 4%

DUB CGI (Stéarinerie Dubois) 5%

DUB OK18 (Stéarinerie Dubois) 1%

MC 30 (Sophim) 1%

Sorbirol L (Cesalpinia) 1%

DUB Wax A (Stéarinerie Dubois) 6.3%

Phase C. Conservative 0.7%

Active principle according to the invention (example 1) 3%

The quantities indicated are given as a percentage of weight.

This emulsified gel has a pH of 6.5.

It can be obtained by the implementation of the following steps:

- Mix A and mix B,

- Heat A and B at 80 ° C,

Emulsifying B in A under a rotor stator emulsifier,

- At 40 ° C add C, homogenize until completely cooled. Example 6 Use of an Active Ingredient of the Invention in a Day Cream

Phase A. Water QSP 100%

Phase B. Lanol 99 (Seppic) 5%

Montanov 202 (Seppic) 3%

Montanov 68 (Seppic) 2%

Sepigel 305 (Seppic) 0.3%

Phase C. Conservative 0.7%

Active principle according to the invention (example 1) 3%

The quantities indicated are given as a percentage of weight.

This emulsified gel has a pH of 6.5.

It can be obtained by the implementation of the following steps:

- Mix A and mix B,

- Heat A and B at 80 ° C,

Emulsifying B in A under a rotor stator emulsifier,

- At 40 ° C add C, homogenize until completely cooled.

EVALUATION OF THE FIRMING EFFECT C "'AN ACTIVE INGREDIENT FROM PLPER NlSRUM

A. In Vitro Studies

These in vitro studies are intended to highlight the effectiveness of an active ingredient according to the invention both on the functionalities and on the synthetic capacities of the reticular dermis.

They were performed on fibroblasts of the reticular dermis obtained from human skin plasty. A particularly suitable sampling and isolation protocol is described in the publication JM SO ELL et al. Journal of cellular physiology, 200: 134-145 (2004). 1. Demonstration of the efficacy of an active ingredient from Piper Niqrum on the functionality of the reticular dermis

i. Study of membership capacities

The objective of this study is to evaluate the effect of an active ingredient derived from Piper Nigrum on the adhesion capacity of reticular fibroblasts to collagen I.

The active ingredient used is that of Example 1.

The study was carried out on normal and photo-aged human reticular fibroblasts obtained using a UVA irradiation.

The operating protocol is as follows.

At 0, human fibroblasts are seeded in a suitable culture medium and the cells are incubated at 37 ° C.

At D3, the cells are treated: the culture medium is removed and replaced with medium containing the active ingredient from Piper nigrum of Example 1 at 0.25 and 0.50% (V / V). The cells are then incubated again at 37 ° C. On D4, the culture medium is removed and replaced with a solution of PBS. The cells are subject or not to UVA.

Normal or photo-aged reticular fibroblasts are trypsinized and inoculated with rat tail collagen I. The cells are then incubated at 37 ° C., then fixed with glutaraldehyde and stained with crystal violet. The visualization of the results is performed on a microscope coupled to an image analysis system. Quantification of the number of cells is performed via a computer program. The higher the staining, the higher the adhesion of the reticular fibroblasts.

The results obtained are shown in the table below: Efficiency / Witness

Optical Density (AU)

(%)

Fibroblasts reticular norir rate

Witness 1.05

Active principle according to the invention 1.11 +6

0.25%

Active ingredient according to the invention 1.13 +8

0.50%

Reticular fibroblasts phot o-old lily

Witness 0.68

Active principle according to the invention 0.79 +15

0.25%

Active principle according to the invention 0.87 +26

0.50%

It is found that the adhesion of photo-aged reticular fibroblasts is significantly reduced by 35% compared to normal reticular fibroblasts.

These results show that the use of an active ingredient derived from Piper nigrum according to the invention, tested at 0.50% makes it possible to increase the adhesion capacity of photo-aged fibroblasts.

ii. Study of contractile power

The objective of this study is to evaluate the ability of an active ingredient derived from Piper Nigrum, to stimulate the contractile power of reticular fibroblasts.

The active ingredient used is that of Example 1.

The operating protocol is as follows. At 0, human reticular fibroblasts are inoculated in a suitable culture medium. The cells are then incubated at 37 ° C.

At D3, the cells are treated: the culture medium is removed and replaced by a medium containing the active principle of Example 1 at 0.25 and 0.50% (V / V) or vitamin C at 0.5 μg / ml. The cells are then incubated at 37 ° C. On D4, the culture medium is removed and replaced with a PBS solution. The cells are subject or not to UVA.

Normal or photo-aged reticular fibroblasts are added to a collagen gel made from rat tail collagen I, containing or not the active ingredient at 0.25% and 0.5% or vitamin C at 0.5 μg / ml. ml.

After 2 hours of incubation at 37 ° C, the gel is peeled off.

At day 7, the diameter of the collagen gels is measured.

The results obtained are shown in the table below:

Contraction capacity

Difference diameter (J4-J7) (cm)

Normal reticular fibroblasts

Witness 1.48

Vitamin C 0.5pg / ml 1.90

Active principle according to the invention 0,50% 1.80

Photo-Aged Reticular Fibroblasts

Witness 0.48

Vitamin C 0.5pg / ml 1.43

Active ingredient according to the invention 0.25% 1.15

Active ingredient according to the invention 0,50% 1.23 It is found that the contractile capacity of photowallic reticular fibroblasts is significantly reduced by 68% compared with normal reticular fibroblasts.

The results obtained show that an active ingredient derived from Piper nigrum according to the invention makes it possible to reduce the diameter of the collagen gels and thus to increase by 156% the contractile power of the reticular fibroblasts.

iii. Study of the power of migration

The purpose of this study is to evaluate the ability of the active principle of the invention to stimulate the migration capacity of the reticular fibroblasts. This study was carried out on normal and photo-aged reticular human fibroblasts obtained using a UVA irradiation.

The operating protocol is described following.

At 0, human reticular fibroblasts are seeded in suitable medium. The cells are then incubated at 37 ° C in an atmosphere containing 5% CO 2.

At J2, two parallel wounds are made crossing the well from left to right.

The culture medium is removed and replaced by a suitable medium. The cells are subject or not to UVA. After irradiation, the culture medium is removed and replaced with medium containing the active ingredient at 0.5 and 2% (V / V).

Photos of the scars are taken every 10 minutes over a period of 48 hours. These images are gathered in a stack of images according to a temporal criterion.

From these sequences, the trajectory of the selected cells is followed. A set of coordinates of these cells is thus obtained as a function of time. The total distance traveled is calculated by cumulating the distance separating each point. The results are as follows

This study shows that the migration of photo-aged reticular fibroblasts is significantly reduced by 25% compared to normal reticular fibroblasts.

Tested at 0.5%, the active ingredient from Piper nigrum significantly increases the migration capacity of photo-aged reticular fibroblasts by 20%.

2. Demonstration of the efficacy of the active ingredient derived from Piper Nigrum on the expression of matrix proteins

The objective of this study is to evaluate the effect of an active ingredient derived from Piper Nigrum according to the invention on its ability to increase the expression of mRNAs encoding fibrillar molecules: collagen I and elastin , the adhesion molecule: α2β1 integrin and the matricellular signaling molecule: CT & F.

The active ingredient used is that of Example 1. The study was carried out by quantitative PCR on normal and photo-aged human reticular fibroblasts obtained using a UVA irradiation. The operating protocol is as follows.

At 0, normal human reticular fibroblasts are inoculated in a suitable culture medium and the cells are then incubated at 37 ° C. At J1, the cells are pre-treated: the culture medium is removed and replaced with medium containing the active principle of Example 1 at 0.5% and 1% (V / V) or TGFp-1 at lng. / ml (reference molecule). The cells are then incubated for 24 hours at 37 ° C.

At D2, the culture medium is removed and replaced with a PBS solution. Irradiation is performed using a UVA lamp.

The PBS solution is removed and replaced with a medium containing the active principle of Example 1 at 0.5% and 1% (V / V) or TGFp-1 at 1 ng / ml. The cells are incubated again.

The cells are recovered for assaying the α2β1 integrin or for assaying collagen I, elastin or CT & F. The cells are recovered and the total RNAs extracted.

The RNAs are reverse transcripts and the complementary DNAs obtained are analyzed by the quantitative PCR technique. The mRNAs of the ribosomal protein S27, an internal reference reference, are also analyzed in parallel with the collagen I, I elastin mRNAs, the α2β1 integrin and the CT & F. Quantification of fluorescence incorporation (SYBR Green) is measured continuously using a thermal cycler. Ct analysis (relative quantization) is performed using software.

The results obtained for each molecule are presented in the tables below: Collégqène I:

Elastin:

Inteqrine α2 (¾1:

CT6F:

It is found that the expressions of the mRNAs of collagen I, elastin, α2β1 integrin and CT & F by photo-aged human fibroblasts are 75%, 49%, 83% and 77%, respectively, compared to normal human fibroblasts.

The results obtained show that an active ingredient from Piper nigrum according to the invention makes it possible to increase the synthesis of collagen I, elastin, α2β1 integrin and CT & F in photo-aged reticular fibroblasts.

B. In Vivo studies

For these trials, a preliminary study assessed the level of cumulative exposure to UV during the lifetime of panelists based on a questionnaire allowing to discriminate two groups of volunteers with a different level of exposure named photo-aged group and non-photo-aged group:

- Group A - non-photo-aged: 19 volunteers, mean age 59, and phototype 3.

- Group B - photo-aged: 19 volunteers of middle age 59 years and phototype 3.

These tests consist of studying, firstly, the impact of aging related to solar exposure on the biomechanical properties of the skin, and then evaluating the effect of an active ingredient derived from Piper nigrum on the improvement of the cutaneous parameters of each skin. of the two groups, and finally to study the effect of the active ingredient on the reduction of existing differences between the two groups on the various parameters relating to the biomechanical properties of the skin. 1. Evaluation of the anti-loosening effect of an active ingredient from Piper Niqrum

At first, the objective of this study is to study the impact on the relaxation of aging skin due to solar exposure.

In a second step, the objective of the study is to investigate the effect of the active ingredient derived from Piper nigrum on the improvement of cutaneous parameters, and thus to study the effect of the active ingredient on the reduction of differences. between the two groups.

The study is performed against placebo. The active ingredient according to the invention which is used is that of Example No. 1 formulated in a composition of Example No. 6.

The evaluation of the parameters related to the relaxation of the face is carried out blindly by a trained expert before and after 56 days of twice-daily application, on the parameters relating to the ptosis of the face and to the firmness.

The evaluation of the ptosis is carried out by visual observation of the expert skin folds formed near the ear. The number of folds formed and their width are indicators of loosening of the skin of the face. In addition, the expert lifts the skin with the index finger and observes how the skin regains its place. A firm skin, not relaxed, has no fold at this point of the face unlike a skin marked and relaxed.

The evaluation of the firmness of the skin is evaluated by the expert by exerting a sustained pressure on the cheek of the volunteer using the fingers index, major and annular. A series of pressure / pressure stops is performed by sweeping an area from the nose to near the ears. These pressure successions make it possible to estimate the resistance of the skin under the fingers. The more the skin is resistant to pressure and therefore to the depression of the fingers, the more it is firm.

The operating protocol of the study is as follows.

Between D-H and Jo, volunteers apply a placebo cream twice a day to the entire face.

At Jo, the volunteers come to the laboratory without applying a product on the face in the morning (no cream or makeup). An evaluation is carried out by the expert.

Between Jo and J55, volunteers perform twice-daily applications of the product and placebo for both groups A and B.

At J56, the volunteers come to the laboratory without having applied a product on the face in the morning (neither cream nor makeup), and the expert carries out a new evaluation.

The results obtained at Jo are presented in the table below:

These first results show that there is a significant difference in the parameters relating to the relaxation of the face evaluated by a trained expert, between the groups of people photovieilli and non photoageilli. A significant decrease in firmness as well as an increase in ptosis is observed to the detriment of the photoweave group. Photoaging has therefore a real impact on the cutaneous parameters and in particular those related to the relaxation of the face.

Furthermore, the results obtained with the active ingredient from Piper Nigrum after 56 days of treatment are presented below as a percentage of variation compared to the results obtained with the placebo:

It is found that under the conditions of this study, in comparison with placebo and after 56 days of twice-daily applications on group A non photoovieilli and group B photovieilli, an active ingredient from Piper nigrum formulated at 3% in emulsion significantly improves the firmness of the skin (+ 14.3%) and limit the ptosis of the face (-8.6%) of people with photoaging skin. The active ingredient according to the invention thus plays specifically on photo-skinned skin with a marked cutaneous relaxation.

The results corresponding to the ptose and firmness parameters evaluated by the expert trained in OJ and J56 are also presented in the following tables: Parameter Ptôse:

Firmness parameter

Under the conditions of this study, after 56 days of twice-daily applications on group B photovieilli, an active ingredient derived from Piper Nigrum according to the invention, formulated at 3% in emulsion, significantly improves the firmness of the skin after clinical evaluation. (+ 17.2%) and decreased ptosis (-14.1%).

The active ingredient derived from Piper Nigrum according to the invention thus allows the group of photowallowed people to find cutaneous characteristics comparable to those of the non-photowalled group, the difference between the two groups being no longer significant at J56. By compensating for the loss of firmness and by limiting the phenomenon of ptosis caused by various environmental factors and more particularly by the UV on the photo-aged group, the active ingredient according to the invention makes it possible to correct the loosening and the sagging of the skins. photo-aged.

2. Evaluation of the effect of an active ingredient derived from Piper Nigrum on the biomechanical properties of the skin

The aim of this study is to study the impact on the biomechanical properties of skin aging related to solar exposure and the effect of an active ingredient from Piper Nigrum on the biomechanical properties of the skin.

The study is performed against placebo. The active ingredient according to the invention used is that of Example No. 1 formulated in a composition of Example No. 6. The evaluation of biomechanical parameters was performed before and after 56 days of twice-daily application using a cheek cutometer ^® . The principle is as follows: the skin is sucked into the orifice of a 6mm diameter probe by a constant depression for a constant duration. Several successive aspirations are made. The depth of penetration of the skin into the probe is measured by means of two optical prisms placed at the opening of the probe. From the curves obtained it is possible to calculate various characteristic parameters of the mechanical properties of the skin.

Among all the parameters calculated from the curves obtained, the present study consists in studying:

the parameter X which corresponds to the tonicity of the skin or elastic retraction

X = (Uf-Ur) = O (1-R7) If -X increases, the tone increases, and

- the parameter O which reflects the firmness of the skin

Uf = RO

If -RO increases, the firmness of the skin increases.

The protocol of the study is as follows.

Between D 4 and Jo, volunteers apply a placebo cream daily on the whole face.

At Jo, the volunteers come to the laboratory without applying a product on the face in the morning (no cream or makeup), and measure the biomechanical properties of the cutometer ^® .

Between Jo and J55, the volunteers apply twice daily in hemi-age product containing the active ingredient according to the invention and placebo for both groups.

At D56, the volunteers come to the laboratory without having applied a product on the face in the morning (neither cream nor makeup), and we measure again the biomechanical properties to the cutometer ^® .

The results obtained at Jo are presented in the table below:

These first results show that there is a significant difference in the biomechanical parameters of the skin, between groups of people "photovieilli" and "non photoageilli" of the same average age. A decrease in the firmness and tone of the skin is observed to the detriment of the group photoweaving. Photoaging therefore has a real impact on the cutaneous parameters related to the firmness and the tonicity of the skin.

It is found that, under the conditions of this study, in comparison with the placebo and after 56 days of twice-daily applications on group A "non photo-photoilli" and group B "photovieilli", the active ingredient from Piper nigrum according to the invention , formulated at 3% in emulsion significantly improves the biomechanical properties of people with photoaging skin.

The active ingredient according to the invention thus plays specifically on skins having altered biomechanical properties characteristic of a greater sagging skin.

The results corresponding to the change in firmness and tonicity parameters measured at the cutometer® at D0 and D56 are also presented in the following tables: Firmness parameter:

Tonicity parameter:

Under the conditions of this study, after 56 days of twice-daily applications on group B photovieilli, the active ingredient according to the invention, formulated at 3% in emulsion, significantly improves the characteristic parameters of the firmness and the tonicity of the skin.

The use of an active ingredient derived from Piper nigrum according to the invention thus allows the group of photowallied persons to recover firmness and tonicity characteristics comparable to those of the non-photowalled group, the difference between the two groups no longer being significant at J56.

Claims

1. Cosmetic active ingredient, characterized in that it is derived from Piper nigrum berries and in that it comprises oligosaccharides with a degree of polymerization of between 2 and 7.

2. Cosmetic active ingredient according to claim 1, characterized in that it comprises at least 10% of oligosaccharides with a degree of polymerization of between 2 and 7, by weight relative to the total content of dry matter.

3. Cosmetic active ingredient according to one of claims 1 or 2, characterized in that the oligosaccharides are in the form of α-glucans and rhamnogalacturonans.

4. Cosmetic active ingredient according to one of the preceding claims, characterized in that it also comprises proteins and uronic acids.

5. Cosmetic active ingredient according to one of the preceding claims, characterized in that it comprises a total sugar content of between 20 and 53% by weight relative to the total content of dry matter.

6. Cosmetic active ingredient according to one of the preceding claims, characterized in that it has at least one of the following characteristics, the percentages being by weight relative to the total content of dry matter:

- a total sugar content of between 20% and 53%,

an ash content of between 28.4 and 42.8%,

a protein content of between 22.7% and 34.1%.

7. Cosmetic active ingredient according to one of the preceding claims, characterized in that it has a solids content of between 17 and 25 g / l and / or a pH between 3.0 and 4.0.

8. Cosmetic active ingredient according to one of the preceding claims, characterized in that it is a hydrolyzate of Piper nigrum berries.

9. Cosmetic active ingredient according to one of the precedingenen claims, characterized in that it does not contain piperine.

10. Use of at least one extract derived from Piper nigrum as a cutaneous firming cosmetic active ingredient.

11. Use of at least one extract derived from Piper nigrum according to claim 10, as a cosmetic active ingredient in a cosmetic composition intended to fight against sagging skin tissues and / or to increase the tone and firmness of the skin. skin.

12. Use of at least one extract derived from Piper nigrum according to any one of claims 10 or 11, as a cosmetic active ingredient in a cosmetic composition intended to combat the loosening of the skin of the face.

13. Use of at least one extract derived from Piper nigrum according to any one of claims 10 to 12, as a cosmetic active ingredient in a cosmetic composition intended to act on the reticular dermis.

14. Use of at least one extract derived from Piper nigrum according to one of claims 10 to 13, as a cosmetic active principle in a cosmetic composition intended to preserve the functionalities and synthetic capacities of the reticular dermis in the face of UV irradiations. .

15. Use of at least one extract derived from Piper nigrum according to one of claims 10 to 14, as a cosmetic active principle in a cosmetic composition intended to increase the adhesion, contraction and migration of the cells of the reticular dermis. .

16. Use of at least one extract derived from Piper nigrum according to any one of claims 10 to 15, as a cosmetic active ingredient inducing the synthesis of collagen I, elastin, α2β1 integrin and / or CT & F in the cells of the reticular dermis.

17. Use of at least one extract derived from Piper nigrum according to one of claims 10 to 16, wherein the extract is an active ingredient according to one of claims 1 to 9.

18. Cosmetic composition for topical application, characterized in that it comprises an active ingredient according to one of claims 1 to 9 present between 0.01 and 20% by total weight of the composition.

19. A cosmetic process for the care of human skin, intended to firm the skin, comprising the topical application of a composition containing an active ingredient derived from Piper nigrum.

20. Cosmetic process for the care of human skin, intended to fight against the collapse of cutaneous tissues, to fight against the loosening of the skin of the face, and / or to strengthen the tone and the firmness of the skin, including the application topical composition containing an active ingredient from Piper nigrum.

21. Cosmetic process for the care of human skin according to one of claims 19 or 20, characterized in that the active ingredient from Piper nigrum is an active ingredient according to one of claims 1 to 9.