WO2021152250A1

WO2021152250A1 - Cosmetic, nutraceutical or dermatological use of a tamarindus indica l. extract and/or a composition comprising it

Info

Publication number: WO2021152250A1
Application number: PCT/FR2021/050135
Authority: WO
Inventors: Vincent Bardey; Florence Henry; Camille MAS; Philippe Moser; Nicolas Pelletier
Original assignee: Basf Beauty Care Solutions France Sas
Priority date: 2020-01-30
Filing date: 2021-01-26
Publication date: 2021-08-05
Also published as: FR3106756A1; FR3106756B1

Abstract

The present invention relates to the cosmetic, nutraceutical and/or pharmaceutical, in particular dermatological, use of a Tamarindus indica L. extract and/or a composition comprising it for maintaining and/or increasing the expression of extracellular matrix molecules of the skin and/or mucosa, the extract not being obtained by using butylene glycol as the extraction solvent.

Description

Cosmetic, nutraceutical or dermatological use of an extract of Tamarindus indica L. and / or of a composition comprising it

TECHNICAL AREA

The present invention relates to the cosmetic, nutraceutical and / or pharmaceutical use, in particular dermatological, of an extract of leaves of Tamarindus indica L. and / or of a composition comprising it for maintaining and / or increasing the expression of molecules of the extracellular matrix of the skin and / or the mucous membranes, the extract not being obtained by using butylene glycol as extraction solvent.

TECHNOLOGICAL BACKGROUND

The structure and properties of the skin change under the effect of complex biological, physical and biomechanical processes, leading to a loss of elasticity, firmness, density of the dermis, especially under the effect of the modification of the molecules of the matrix extracellular.

The extracellular matrix of the skin and mucous membranes is made up of the association of three types of molecules: fibers: collagen and elastin, glycoproteins: less abundant than fibers but have a more important role in cell adhesion, such as fibronectin and laminin, highly hydrated polysaccharides constituting a gel filling the matrix. This matrix plays a major role in maintaining the structure and properties of the skin, in particular the elasticity, firmness and density of the dermis. Since these molecules are synthesized in particular by fibroblasts and keratinocytes, the renewal of the latter is essential for maintaining the biomechanical properties of the skin.

Elastin is a protein that makes up elastic fibers by associating with other molecules such as fibrillins and MAGPs (Microfibrillar Associated Glycoproteins).

Elastin is synthesized in the form of soluble tropoelastin which acquires its physicochemical properties (insolubility, elasticity) after its intra- and intermolecular crosslinking by a lysyl oxidase (LOX) and its deposition on the microfibrils. The elastic fibers are responsible for the elastic property of the organs that contain them (vessels, pulmonary parenchyma, elastic cartilages, skin, etc.). The name elastin is reserved for the protein forming the amorphous portion of elastic fibers and giving them their elasticity.

The aging of the skin and mucous membranes is associated with a modification of the fibrillar networks, in particular that of the elastic fibers which degrade and no longer reform correctly.

In the skin, functional elastic fibers are formed by fibroblasts in the dermis, but certain components necessary for the development of elastic fibers have also been found in the cells of the epidermis. The rate of renewal of elastic fibers is very low in adulthood, although the overall level of elastin in the skin may increase. In newborns, not all micro fibrils are completely covered with elastin, and become so around puberty. From the age of 40, we see the appearance of inclusions on the fibers, more frequent in women, then the fragmentation of the elastic fibers and their disappearance under the dermo-epidermal junction (DEJ). This fractionation and / or this disappearance under the DEJ results in the loss of elasticity of the skin and the formation of wrinkles. Synthesis of non-functional elastic fibers is observed during photoaging, but this increase is accompanied by the accelerated loss of elastic fibers under the DEJ.

Collagen is a constituent protein of the extracellular matrix (ECM) present in large quantities in the tissues of vertebrates. This is a large family comprising 29 different types.

Among the different types of collagen, type I collagen is found in particular in many human tissues such as tendons, ligaments, cornea and skin, located more precisely in the dermis. Type I collagen is the major collagen in the skin.

Fibrillar collagen is formed from procollagen fibers, then assembled into a network, then stabilized by crosslinking. It is collagen that gives tissues their mechanical strength, and as a result, helps maintain their firmness.

As the skin ages, the level of collagen generally decreases, resulting in a loss of firmness. This phenomenon is all the more pronounced since the skin is, among other things, subjected to UV rays. We talk about photobiological aging.

Several mechanisms are involved in the decrease in collagen levels during tissue aging: enzymes called collagenases are responsible for the degradation of collagen proteins. In parallel, a non-enzymatic glycation mechanism at the origin of the formation of glycated proteins called AGEs (Advanced Glycation End Products), in the extracellular matrix, participates in a decrease in the functionality of said matrix, and therefore of the skin. .

The molecules of the extracellular matrix are therefore the subject of numerous studies and innovations in the field of cosmetics for the development and improvement of cosmetic active ingredients aimed at maintaining the biomechanical properties of the skin.

Tamarindus indicaL. Commonly called tamarind or tamarind is a 12-15 m tree that can reach 30 m in height, nowadays distributed in a large number of warm regions of the globe. It is most likely originally from tropical Africa (especially Burkina Faso) from where it spread in the past in all the hot parts of the globe. The main production areas of T. indica are in Asia (India, Bangladesh, Sri Lanka, Thailand, Vietnam and Indonesia) and America (Mexico, Belize, Costa Rica and Brazil) Tamarindus indica is mainly used for its fruits, especially the pulp, which has a wide variety of domestic and industrial uses. The seed is commercially available as a food additive to improve the viscosity and texture of processed foods. The leaves, flowers and immature pods are also edible. The leaves and flowers are used in making curries, salads, stews and soups in many countries, especially in times of food shortage but also as a bite in tincture.

Tamarindus indica leaves are also used for various applications in traditional medicine, including in wound healing, treatment of inflammation, treatment of snakebites, diarrhea and liver pain.

Various studies carried out on extracts from T. indica leaves have demonstrated the antioxidant properties of methanolic leaf extracts, the antiglycation properties of ethanolic extracts of a residue obtained after extraction with ethyl acetate from leaves of Tamarindus indica , the anti-bacterial and antifungal activities of aqueous, acetonic and ethanolic extracts of bark of stems and leaves of Tamarindus indica of Niger origin and anti-inflammatory and antinociceptive activities of a hydro-ethanolic extract (ethanol 95% -water 5%) of Tamarindus indica leaves of Indian origin. Document KR20130032995 thus discloses the use as an anti-inflammatory agent of an extract of Tamarind.

Finally, patent application US2012189725 discloses the inhibitory effect of the elastase, TNF-α and MMPI activities of an extract of Tamarindus indica leaf obtained with butylene glycol as an extraction solvent. Another extract of Tamarind pericarp exhibiting anti-elastase and anti-collagenase activities has been described in application US2008 / 286387. It is obtained by methanol extraction of the polyphenols contained in the pericarp of Tamarind.

SUMMARY OF THE INVENTION

Thus, to the knowledge of the Applicant, no prior art discloses or suggests the cosmetic, nutraceutical and / or pharmaceutical, in particular dermatological use of an extract of Tamarindus indica leaves and / or of a composition comprising it to maintain and / or increase the expression of molecules of the extracellular matrix of the skin, mucous membranes, and / or to maintain and / or increase the expression of collagen fibers and / or elastic fibers of healthy skin and / or healthy mucous membranes, and / or to maintain and / or increase the biomechanical properties of healthy skin and / or healthy mucous membranes, and / or to maintain and / or increase the proliferation of fibroblasts of the skin and / or mucous membranes, and / or or to maintain and / or increase the differentiation of the keratinocytes of the skin and / or of the mucous membranes, the extract not being obtained using butylene glycol as extraction solvent. The first subject of the present invention is the cosmetic and / or nutraceutical use of an extract of Tamarindus indica leaves to maintain and / or increase the expression of molecules of the extracellular matrix of healthy skin and / or healthy mucous membranes, the extract not being obtained by using butylene glycol as extraction solvent.

It also relates to a cosmetic care process characterized in that it comprises the topical application to at least one area of healthy skin and / or healthy mucosa of an extract of Tamarindus indica as defined above. , or of a cosmetic composition comprising it, to maintain and / or increase the expression of molecules of the extracellular matrix of healthy skin, of healthy mucous membranes, and / or to maintain and / or increase the expression of collagen fibers and / or elastic fibers of healthy skin and / or healthy mucous membranes, and / or to maintain and / or increase the biomechanical properties of healthy skin and / or healthy mucous membranes, preferably firmness and / or elasticity.

A subject of the invention is also an extract of Tamarindus indica as defined above, or of a dermatological composition comprising it, for its use in the treatment and / or prevention of pathologies of the skin and / or of the mucous membranes involving loss of expression of molecules of the extracellular matrix of the skin and / or mucous membranes, and / or loss of expression of collagen fibers and / or elastic fibers of the skin and / or mucous membranes, and / or a loss of the biomechanical properties of the skin and / or of the mucous membranes, in particular a loss of firmness of the skin and / or of the mucous membranes, and / or a loss of the proliferation of fibroblasts of the skin and / or of the mucous membranes, and / or loss of keratinocyte differentiation of the skin and / or mucous membranes.

DETAILED DESCRIPTION

For the purposes of the present invention, the term “cosmetic use and / or composition” is understood to mean a non-pharmaceutical use and / or composition, that is to say which is not intended for therapeutic use and is applied to a non-pharmaceutical use. so-called healthy part of the body, in particular in a so-called healthy area of the skin and / or mucous membranes.

For the purposes of the present invention, the term "nutraceutical use and / or nutraceutical composition" means a use and / or a composition for non-pharmaceutical oral administration, that is to say which does not require therapeutic treatment.

For the purposes of the present invention, the term “healthy skin”, “healthy mucosa” means an area of skin or mucosa to which the extract according to the invention is applied and said to be “non-pathological” by a. dermatologist, that is to say not presenting any infection, scar, disease or skin condition such as candidiasis, impetigo, psoriasis, eczema, acne or dermatitis, or wounds or injuries.

According to the invention, the term “mucosa (s)” denotes the ocular mucosa, the vaginal mucosa, the urogenital mucosa and / or the oral mucosa, in particular the buccal, labial and / or gingival mucosa, preferably the ocular mucosa and / or buccal, and more preferably, the labial and / or ocular mucosa.

For the purposes of the present invention, the term “maintaining and / or increasing the expression of the molecules of the extracellular matrix” is understood to mean preventing a decrease and / or increasing the level of gene expression, that is to say mRNAs. (Messenger RNA), and / or protein synthesis of molecules of the extracellular matrix of healthy skin and / or healthy mucous membranes, treated with the extract according to the invention, relative to the level of gene expression and / or of protein synthesis detected in the absence of the extract according to the invention. Preferably, it involves maintaining and / or increasing the gene expression of molecules of the extracellular matrix.

For the purposes of the present invention, the term “molecules of the extracellular matrix” means all the fibers and / or glycoproteins of the extracellular matrix of healthy skin and / or healthy mucous membranes. Preferably, they are collagen fibers and / or elastic fibers of the extracellular matrix of healthy skin and / or healthy mucous membranes.

Thus, the present invention more particularly aims to maintain and / or increase the expression of collagen fibers and / or elastic fibers of the extracellular matrix of healthy skin and / or healthy mucous membranes.

For the purposes of the present invention, the term “maintaining and / or increasing the expression of collagen fibers and / or elastic fibers of healthy skin and / or healthy mucous membranes” is understood to mean preventing a decrease and / or increasing the level of 'gene expression, that is to say the expression of mRNAs (messenger RNAs), and / or protein synthesis of collagen fibers and / or elastic fibers of healthy skin and / or healthy mucous membranes treated with the extract according to the invention, relative to the gene expression and / or protein synthesis detected in the absence of the extract according to the invention.

In particular, it is an increase in the level of gene expression and / or of the protein synthesis of the collagen fibers of at least 40%, preferably of at least 200% and even more preferably of at least 400%, relative to the level of gene and / or protein expression of collagen fibers and / or elastic fibers measured in the absence of the extract of Tamarindus indica according to the invention. In particular, it is an increase in the level of gene expression and / or protein synthesis of collagen fibers and / or elastic fibers of at least 20%, preferably at least 40% and again more preferably at least 50%, relative to the level of gene and / or protein expression of collagen fibers and / or elastic fibers measured in the absence of the extract of Tamarindus indica according to the invention.

Preferably, it is about maintaining and / or increasing the protein synthesis of collagen fibers and / or elastic fibers.

For the purposes of the present invention, the term “collagen” is understood to mean the type I, III, IV, V, VI, VII, XII, XIII, XIV, XVI, XVII, XXIV, XXIX type collagen proteins present in the skin. healthy and / or healthy mucous membranes. According to an advantageous embodiment, the collagen fibers are chosen from type I, III, IV, V and / or XIV collagen fibers, preferably type I, IV and / or V collagen fibers, even more preferably type I, IV and / or V collagen fibers. type I collagen.

For the purposes of the present invention, the term “type I collagen” protein is understood to mean the collagen protein present in the skin, the cornea, the tendons, the ligaments, the bones, more preferably in the skin. Thus, in a preferred embodiment of the invention, the extract of Tamarindus indica leaves according to the invention is therefore used to maintain and / or increase the protein expression of type I collagen in healthy skin and / or healthy mucous membranes.

According to an advantageous embodiment, it is the increase in the gene expression of the collagen fibers of one or more genes chosen from LAMB1 (encoding the Laminin subunit b 1 protein), LAMG1 (encoding the Laminin subunit protein g 1), NID-1 (encoding the Nidogen 1 protein), COL7A1 (encoding the Collagen Type VII a 1 Chain protein), BGN (encoding the Biglycan protein), SP ARC (encoding the Secreted protein acidic & protein) cystein rich), COL1A2 (encoding the Col I 2 chain protein), COL3A1 (encoding the 1 chain Col III protein), COL1A (encoding the 1 chain Col I protein), COL1A2 (encoding the Col protein I a 2 chain), COL3A1 (encoding the 1 chain Col III protein), COL4A1 (encoding the 1 chain Collagen Type IV protein), COL4A2 (encoding the 2 chain Collagen Type IV protein), COL5A1 (encoding for Col V al chain protein), COL5A2 (coding for Col V a 2 chain protein), COL14A1 (coding for Col XIV 1 chain protein), P4HA1 (c odant for Prolyl 4-Hydroxylase Subunite al protein), P4HA2 (encoding for Prolyl 4-Hydroxylase Subunite a 2 protein), P4HA3 (encoding for Prolyl 4-Hydroxylase Subunite a 3 protein), P4HB (encoding for Prolyl 4 protein - Hydroxylase Subunite b). According to an advantageous embodiment, it is an increase in the level of gene expression of the elastic fibers of one or more genes chosen from Elastin, Fibulin 1, Fibrilin 1 and / or Microfibril associated protein 2.

According to an advantageous embodiment of the invention, it is an increase relative to the level of gene and / or protein expression of collagen measured in dermal fibroblast cells cultured in vitro in the absence of the extract of Tamarindus indica according to the invention.

In a preferred embodiment of the invention, it is an increase in the level of protein expression of type I collagen of at least 50%, preferably of at least 200% and even more preferably of at least 400% relative to the level of protein expression measured in dermal fibroblast cells cultured in vitro in the absence of the extract according to the protocol as described for example in Example 4.

Advantageously, the measurement of the increase in protein expression is carried out by in vitro measurement, preferably after immunostaining, for example according to the method as presented in Example 4 and / or 5.

For the purposes of the present invention, the term “immunostaining” is understood to mean the technique consisting in fixing an antibody specific for a given collagen protein, preferably the type I collagen protein, on said protein, with a view to being revealed by microscopic observation. Advantageously, the revelation is done by confocal microscopy.

According to another advantageous embodiment, the measurement of the increase in protein expression is carried out by in vitro measurement, preferably after RT-qPCR, for example according to the method as presented in Example 6.

In a preferred embodiment, the extract according to the invention is used to maintain and / or increase the biomechanical properties of healthy skin and / or healthy mucous membranes, preferably firmness and / or elasticity and / or density of healthy skin and / or healthy mucous membranes, more preferably the firmness and / or elasticity of healthy skin and / or healthy mucous membranes.

For the purposes of the present invention, the term “biomechanical properties of healthy skin and / or healthy mucous membranes” means firmness and / or elasticity and / or density and / or resistance to compression and / or resistance to compression. stretch and / or flexibility and / or extensibility and / or the ability to resist deformation of healthy skin and / or healthy mucous membranes. Preferably it is the firmness and / or elasticity and / or density of healthy skin and / or healthy mucous membranes, in particular of the healthy dermis, more preferably it is the firmness and / or elasticity of healthy skin and / or healthy mucous membranes, in particular of the healthy dermis.

The biomechanical properties of healthy skin and / or healthy mucous membranes can be studied by measurement techniques known to those skilled in the art, in particular by traction, by torsion, suction, indentation, levatometry, ballistometry, by high-resolution ultrasound scanner or elastography, preferably by indentation, by high resolution ultrasound scanner.

For the purposes of the present invention, the term “maintaining and / or increasing the biomechanical properties of healthy skin and / or healthy mucous membranes” is understood to prevent a reduction and / or increase in the biomechanical properties of healthy skin and / or healthy mucous membranes. , preferably the firmness and / or elasticity of healthy skin and / or healthy mucous membranes, in particular healthy dermis, treated with the extract according to the invention with respect to the biomechanical properties of healthy skin and / or mucous membranes healthy, measured in the absence of the extract according to the invention.

Preferably, this is an increase of at least 5%, preferably 10%, in the biomechanical properties of the healthy skin and / or of the healthy mucous membranes treated with the extract of Tamarindus indica according to the invention compared to the biomechanical properties. healthy skin and / or healthy mucous membranes measured in the absence of the extract of Tamarindus indica according to the invention.

In particular, maintaining and / or increasing the biomechanical properties of healthy skin makes it possible to limit and / or reduce the sagging of healthy skin of the face, in particular in the cheeks and / or the angular zone of the face ( commonly referred to as roundness of the face or V of the face).

For the purposes of the present invention, from a cosmetic point of view, the expression “maintain and / or increase the firmness of healthy skin and / or healthy mucous membranes”, prevent the decrease and / or increase for aesthetic purposes, firmness of healthy skin and / or healthy mucous membranes, in particular those which have lost firmness, in particular under the effect of intrinsic factors. The intrinsic factors can be tissue aging of the skin and / or mucous membranes, i.e. chrono-induced aging, which is found for example in so-called mature skin, i.e. skins of persons over 40 years of age. It may be cellular stress, non-pathological physiological variations such as a change in diet, hormonal variations, in particular during puberty, pregnancy, menopause and / or andropause. This loss of firmness can also appear under the effect of extrinsic factors such as aggressive environmental agents such as UV radiation, pollution, smoke, tobacco, toxins, climatic and / or mechanical aggressions. The increase in firmness can be measured according to conventional methods, in particular by in vivo measurement by the fringe projection method using a Cutometer®, a DensiScore®, a torquemeter or even a DynaSKIN associated with a DermaTOP.

For the purposes of the present invention, from a cosmetic point of view, the term “maintaining and / or increasing the elasticity of healthy skin and / or healthy mucous membranes” is understood to mean preventing the reduction and / or causing the increase at aesthetic purposes, the elasticity of healthy skin and / or healthy mucous membranes which have lost elasticity in particular under the effect of intrinsic factors such as tissue aging of the skin and / or mucous membranes, that is, i.e. chrono-induced aging, which is found for example in so-called mature skin i.e. the skin of people over 40 years old, cellular stress, non-pathological physiological variations such as change in diet, hormonal variations, especially during puberty, pregnancy, menopause and / or andropause. This loss of elasticity can also appear under the effect of extrinsic factors such as aggressive environmental agents such as UV radiation, pollution, smoke, tobacco, toxins, climatic and / or mechanical aggressions. .

The increase in elasticity can be measured according to conventional methods, in particular by in vivo measurement using a Cutometer®, a torquemeter or even a DynaSKIN.

The extract of Tamarindus indica according to the invention can be used to maintain and / or increase the proliferation of fibroblasts in healthy skin and / or in healthy mucous membranes, and / or to maintain and / or increase the differentiation of keratinocytes in the skin. healthy and / or healthy mucous membranes.

Unless otherwise indicated, the term "fibroblasts" and / or "keratinocytes" means "healthy" fibroblasts and / or "healthy" keratinocytes, that is to say not showing any pathology.

For the purposes of the present invention, the term “increasing the proliferation of fibroblasts” is understood to mean an increase in the proliferation of fibroblasts of at least 5%, preferably of at least 10% and even more preferably of at least 20%, relative to the level of fibroblast proliferation measured in the absence of the extract of T. indica according to the invention.

The measurement of the proliferation of fibroblasts can be carried out by any standard method known to those skilled in the art. It can be carried out by assaying DNA, by incorporation of fluorescent nucleic bases, by incorporation of tritiated thymidine, by labeling of proteins specific to proliferating cells with Ki67, by counting cells with a flow cytometer and / or by measurement. density optical. Advantageously, it is carried out by in vitro measurement, by measurement of the optical density, for example according to the method as presented in Example 2.

For the purposes of the present invention, the term “increasing the differentiation of keratinocytes” is understood to mean an increase in the differentiation of keratinocytes of at least 5%, preferably of at least 10% and even more preferably of at least 20%, relative to the level of differentiation of the keratinocytes measured in the absence of the extract of T. indica according to the invention.

The measurement of the differentiation of the keratinocytes can be carried out by any conventional methods known to those skilled in the art, in particular by assaying the involucrine and / or the filaggrin. Advantageously, it is carried out by in vitro measurement, by measurement of the synthesis of involucrine, for example according to the method as presented in Example 3.

For the purposes of the present invention, the extract of Tamarindus indica according to the invention is not obtained using butylene glycol as extraction solvent.

According to a preferred embodiment, the extract of Tamarindus indica leaves according to the invention is useful for maintaining and / or increasing the expression of molecules of the extracellular matrix of healthy skin, of healthy mucous membranes, and / or for maintaining and / or or increase the expression of collagen fibers and / or elastic fibers of healthy skin and / or healthy mucous membranes, and / or to maintain and / or increase the biomechanical properties of healthy skin and / or healthy mucous membranes, preferably firmness and / or elasticity and / or to maintain and / or increase the proliferation of fibroblasts in healthy skin and / or healthy mucous membranes, and / or to maintain and / or increase the differentiation of keratinocytes in healthy skin and / or healthy mucous membranes.

Even more preferably, the extract of Tamarindus indica leaves according to the invention is useful for increasing the expression of molecules of the extracellular matrix of healthy skin, of healthy mucous membranes, and / or for increasing the expression of collagen fibers and / or elastic fibers of healthy skin and / or healthy mucous membranes, and / or to increase the biomechanical properties of healthy skin and / or healthy mucous membranes, preferably firmness and / or elasticity and / or to increase the proliferation of fibroblasts from healthy skin and / or healthy mucous membranes, and / or to increase the differentiation of keratinocytes from healthy skin and / or healthy mucous membranes, more preferably to increase firmness. The extract according to the invention can be obtained by any method of extracting plants known to a person skilled in the art. The extraction can be carried out in particular by maceration or by other processes such as leaching, decoction, extraction under reflux, percolation, flash-expansion, extraction assisted by enzymes, extrusion, extraction by means of ultrasound and / or microwave. Preferably, the extraction is carried out by maceration.

The leaves are preferably dried and / or crushed before extraction.

The extract according to the invention may in particular be obtained by extraction in a solvent or mixture of solvents, preferably a polar protic solvent with the exception of butylene glycol, and advantageously in water, an alcohol, a glycol, a polyol. , a water / alcohol, water / glycol or water / polyol mixture (such as water mixed with ethanol, glycerol and / or other glycols such as xylitol and / or propanediol, etc.) from 99/1 to 1/99 (w / w), advantageously in water as the sole solvent.

For the purposes of the present invention, the term “protic polar solvent” is understood to mean a solvent having a dipole moment and at least one hydrogen capable of intervening in hydrogen bonds.

In a preferred embodiment, the extract is obtained by aqueous extraction. For the purposes of the present invention, the term “extract obtained by aqueous extraction” is understood to mean any extract obtained by extraction with an aqueous solution, with the exception of solutions comprising butylene glycol, containing more than 60% by weight, advantageously at least 70%. % by weight, in particular at least 80% by weight, more particularly at least 90% by weight, in particular at least 95% by weight, of water relative to the total weight of the aqueous solution, even more advantageously not containing no glycol and in particular not containing alcohol, preferably containing only water. Preferably, the extract according to the invention is not obtained by extraction with an ethyl acetate solvent.

In an alternative embodiment, the extraction could be carried out in the presence of a solvent of C ₆ -Ci ₆ dialkyl carbonate type. By way of example of this embodiment, mention may be made of obtaining an extract according to the invention by adding the part of Tamarindus indica to be extracted, preferably from leaves, to the extraction solvent containing the dialkyl carbonate, preferably chosen from dioctyl carbonate and diethylhexyl carbonate with stirring for 2 hours at 80 ° C.

In another alternative embodiment of the invention, the extraction can be carried out in the presence of a nonionic surfactant, preferably chosen from lauryl glucoside marketed under the name Plantacare® 1200UP by BASF or else caprylyl / capryl glucoside (Plantacare® 810 UP), preferably caprylyl / capryl glucoside (Plantacare® 810 UP). The concentration by weight of the nonionic surfactant may be between 0.5% and 5%, advantageously between 0.5 and 1%, again advantageously it will be 1% by weight relative to the total weight of the extracted and solvent mixture.

According to another embodiment, the extraction can be carried out with a solvent comprising coconut water. According to this embodiment, the extract according to the invention can be obtained by adding the part of Tamarindus indica to be extracted, preferably from leaves (leaf / solvent ratio = 1/10) with stirring with the extraction solvent (a mixture of water, coconut water) whose pH has been adjusted to 3 or 4, for 1 hour at 80 ° C in a closed environment to avoid evaporation of the solvent. The extract is then cooled, centrifuged and filtered at 0.45 µm.

In another alternative embodiment of the invention, the extract can be obtained by extraction under supercritical conditions (CO2) in the presence of a co-solvent. Advantageously, the co-solvent will be ethanol. In another alternative embodiment of the invention, the extraction can be carried out by aqueous extraction under subcritical conditions.

The expression “subcritical conditions” extraction is understood to mean extraction in the presence of water, under conditions of temperature above 100 ° C. and pressure below 22.1 MPa (221 bars), such that the water remains at the temperature. liquid state but has a lower viscosity and surface tension than water at room temperature, increasing its dielectric constant.

Thus, the extraction pressure will for example be between 10 MPa (100 bars) and 25 MPa (250 bars), preferably between 15 and 22.1 MPa (150 and 221 bars).

In general, the extraction can be carried out from dry or fresh material, advantageously dry, in an amount of 0.1% to 20% by weight, advantageously from 1% to 10%, very advantageously from 5% to 10% , and even more advantageously 10%, by weight relative to the total weight of the material and of the extraction solvent.

The extraction can be carried out at a temperature ranging from 4 ° C to 300 ° C, including room temperature, that is to say a temperature of 20 ° C. In a preferred embodiment of the invention, the extraction will be carried out at a temperature of 60 ° C to 90 ° C, preferably from 70 ° C to 85 ° C, more preferably at a temperature of 80 ° C.

In an alternative embodiment of the invention, the extraction will be carried out at a temperature of 4 ° C to 25 ° C, again preferably from 4 ° C to 20 ° C, again advantageously at room temperature, that is to say - say at 20 ° C.

In yet another alternative embodiment of the invention, the extraction will be carried out in water under subcritical conditions, at a temperature ranging from 100 ° C to 300 ° C, advantageously from 120 ° C to 250 ° C, again advantageously between 140 ° C and 200 ° C. The extraction can be carried out at a single given temperature or at successive increasing temperatures. In an advantageous embodiment of the invention, the extraction will be carried out at a single temperature of 160 ° C. In an alternative mode, it will be conducted according to a gradient of three increasing temperatures between 100 ° C and 200 ° C, such as 120 ° C, 140 ° C then 160 ° C or 110 ° C, 130 ° C then 150 ° C , or alternatively 120 ° C, 145 ° C then 170 ° C. The extraction can be carried out during a period ranging from 1 hour to 20 hours, preferably during a period ranging from 2 hours to 16 hours. Again preferably, said extraction is carried out during a period of 1 hour.

The extract of Tamarindus indica leaves can be obtained by extracting a quantity of dry or fresh material, advantageously dry, in an amount of 0.1% to 20% by weight, advantageously from 1% to 10%, very advantageously of 5% to 10%, and even more advantageously 10%, by weight relative to the total weight of the material and the extraction solvent by weight of dry leaf material relative to the total weight of the mixture consisting of the solvent and the sheets (p / p). Preferably, the extract is obtained by extracting an amount of 10% by weight of dry matter from the leaves, relative to the total weight of the mixture consisting of the solvent, preferably water, and the leaves (w / w).

The extracts obtained are then preferably centrifuged and / or filtered and / or distilled, preferably centrifuged, in order to recover the active soluble fraction (crude extract). Additional decolorization and / or deodorization and / or fractionation and / or purification steps can be carried out on the extract at any stage of the extraction and according to techniques known to those skilled in the art. The extract according to the invention can then be concentrated by evaporation of the solvent, by use of membrane technologies or dried, for example by lyophilization, zeodration or by atomization.

In a particular embodiment of the invention, in particular for its use in dermatology, the extract of Tamarindus indica leaves obtained according to the invention can be sterilized.

The extract according to the invention is preferably obtained according to one of the embodiments described below:

- In a first embodiment of the invention, the extract is obtained by extraction in water as the sole solvent of an amount of 10% (w / w) of crushed leaves relative to the total weight of the solvent and of the material, with stirring and for a period of one hour at a temperature of 80 ° C. After cooling, the mixture is centrifuged and the extract is filtered. The extract is obtained in liquid form under the conditions described in Example la).

- In a second embodiment of the invention, the extract prepared according to the preceding procedure is diluted with glycerin, so as to obtain a final glycerin concentration of 80% (v / v) by volume of glycerin by relative to the total volume of glycerin and extract. The extract is obtained in liquid form under the conditions described in Example lb).

- In a 3 ^rd embodiment of the invention, the extract is obtained by maceration in water as a single solvent of an amount of 10% (w / w) by weight of crushed leaves relative to the total weight of solvent and the material, with stirring and for a period of 16 hours at a temperature of 4 ° C. The mixture is centrifuged and the extract is then filtered. The extract is then diluted so as to obtain a final glycerin concentration of 80% (v / v) by volume of glycerin relative to the total volume of glycerin and of the extract. The extract is obtained in liquid form under the conditions described in example 1c).

- In a 4 ^th embodiment, the extract is obtained by extraction with stirring and under reflux of an amount of 10% (w / w) by weight of crushed leaves relative to the total weight of the material and the solvent, in an ethanol: water mixture (70:30; v / v) at a temperature of 80 ° C for a period of one hour. The suspension is filtered then the hydroalcoholic extract recovered. The alcoholic phase is evaporated in vacuo and the residual aqueous phase obtained is centrifuged and filtered. This aqueous phase is atomized in the presence of maltodextrin, in a final amount of maltodextrin of 80% (w / w) by weight relative to the total weight of the extract and of maltodextrin in solid form, under the conditions described in example ld ).

- Into a ^5-th embodiment of the invention, the extract is obtained by extraction in water as a single solvent of an amount of 20% (w / w) by weight of crushed leaves with respect to the weight of the solvent and the material, with stirring and for a period of one hour at a temperature of 80 ° C. The crude extract was centrifuged, then filtered and then atomized in the presence of maltodextrin, in a final quantity of maltodextrin of 80% (w / w) by weight relative to the total weight of the final extract in solid form, under the conditions described. in the example).

- in a 6 ^am embodiment of the invention, the extract is obtained by aqueous extraction in subcritical conditions in the water as the sole solvent of total sheets. The extraction was carried out at a flow rate of 5 ml / min and a solvent / Tamarindus indica leaf ratio of 25 ml / g at a temperature of 120 ° C, and a pressure sufficient for the water to be in its liquid state. , i.e. 16 bars. The extract is then dried and ground to obtain a fine powder under the conditions described in example 1f).

- In a 7 ^th embodiment of the invention, the extract is obtained by aqueous extraction in subcritical conditions in the water as the sole solvent of total sheets.

The extraction was carried out at a flow rate of 5 ml / min and a solvent / Tamarindus indica leaf ratio of 25 ml / g at a temperature of 140 ° C and a pressure sufficient for the water to be in its liquid state, i.e. 18 bars. The extract is then dried and ground to obtain a fine powder under the conditions described in Example 1g).

According to the invention, the extract of Tamarindus indica according to the invention can be used alone, in the form of active ingredient and / or in a cosmetic and / or nutraceutical, and / or pharmaceutical composition, in particular dermatological, preferably intended for topical application and / or oral route, more preferably for topical application.

When it is used alone in the form of active ingredient, the extract according to the invention is preferably soluble and / or diluted in a solvent, in particular polar, such as water, an alcohol, a polyol, a glycol such as pentylene glycol and / or hexylene glycol and / or caprylyl glycol, or one of their mixtures, preferably a hydroglycolic or hydroalcoholic mixture, more preferably comprising a glycol chosen from hexylene glycol, caprylyl glycol and their mixtures.

Preferably, the active ingredient is not diluted in butylene glycol.

Advantageously, the extract according to the invention is soluble and / or solubilized in an aqueous solution comprising glycerol, advantageously in a solution comprising at least 80% (w / w) by weight of glycerol relative to the total weight of the aqueous solution .

Alternatively, the extract obtained is diluted and / or soluble in an aqueous solution comprising caprylyl glycol, in particular comprising between 0.01 and 5% (w / w) by weight of caprylyl glycol, preferably between 0.1 and 1% (w / w) by weight of caprylyl glycol, relative to the total weight of the aqueous solution.

In another embodiment, the extract according to the invention can be incorporated into a cosmetic and / or nutraceutical composition comprising at least one cosmetically acceptable and / or nutraceutical acceptable excipient. Preferably, it is incorporated into a cosmetic composition comprising at least one cosmetically acceptable excipient.

For the purposes of the present invention, the term “cosmetically acceptable” excipient is understood to mean a topically acceptable compound and / or solvent, that is to say not inducing an allergic response on contact with the skin, including the human scalp, and / or mucous membranes, non-toxic, non-unstable, chemically.

In a preferred embodiment of the invention, the extract according to the invention is present in the cosmetic and / or dermatological composition in a content of between 1x10 ⁴ % and 10% (v / v) by volume, preferably between 1x10 ⁴ % and 5% (v / v) by volume, again advantageously between 1 × 10 ³ % and 3% (v / v) by volume, more preferably between 0.1% and 1% (v / v) by volume, relative to to the total volume of the composition, said composition further comprising at least one cosmetically acceptable excipient.

The cosmetic and / or dermatological composition of the invention can be chosen from an aqueous or oily solution, an aqueous cream or gel or an oily gel, in particular a shower gel, a shampoo; a milk ; an emulsion, a microemulsion or a nanoemulsion, in particular oil-in-water or water-in-oil or multiple or silicone; a mask; a serum; a lotion; liquid soap; a dermatological bar; an ointment ; a foam; a patch; an anhydrous product, preferably liquid, pasty or solid, for example in the form of makeup powders, sticks or sticks, in particular in the form of lipstick.

It can also be a makeup product or a makeup removal product.

Advantageously, the extract or the composition of the invention is intended to be applied to all or part of the body and / or of the face and / or of the scalp, preferably the legs, thighs, arms, stomach, the décolleté, the neck, the armpits, the lips, more preferably all or part of the face, and preferentially the cheeks, the forehead, the chin, the lips, the contour of the eyes.

Advantageously, the extract and / or the composition comprising it is intended to be applied to an area of healthy skin exhibiting sagging and / or an area of sagging healthy skin and / or an area of healthy skin lacking in tone.

Thus, the cosmetic composition is advantageously intended for topical application to healthy skin and / or healthy mucous membranes.

Preferably, the extract according to the invention is particularly suitable for the formulation of a so-called neutral and gentle composition for respecting the sebaceous gland, in particular the skin including the scalp and / or mucous membranes.

Alternatively, the extract according to the invention can be in all the dosage forms conventionally used for oral application, in particular in the form of a nutraceutical active ingredient and / or of a nutraceutical composition.

The compositions according to the invention may contain any suitable solvent and / or any suitable vehicle and / or any suitable excipient, optionally in combination with other compounds of interest.

Therefore, for these compositions, the excipient contains for example at least one compound chosen from the group comprising preservatives, emollients, emulsifiers, surfactants, moisturizers, thickeners, conditioners, mattifying agents, stabilizers. , antioxidants, texturing agents, shine agents, film-forming agents, solubilizers, pigments, dyes, perfumes and sun filters. These excipients are preferably chosen from the group consisting of amino acids and their derivatives, polyglycerols, esters, polymers and cellulose derivatives, lanolin derivatives, phospholipids, lactoferrins, lactoperoxidases, stabilizers based on sucrose, vitamins E and its derivatives, natural and synthetic waxes, vegetable oils, triglycerides, unsaponifiables, phytosterols, vegetable esters, silicones and its derivatives, protein hydrolysates, Jojoba oil and its derivatives, lipo / water-soluble esters, betaines, aminoxides, plant extracts, sucrose esters, titanium dioxides, glycines, and parabens, and more preferably from the group consisting of butylene glycol, steareth-2, steareth-21, glycol-15 stearyl ether, cetearyl alcohol, phenoxyethanol, methylparaben, ethylparaben, propylparaben, butylparaben, butylene glycol, natural tocopherols, glycerin, dihydroxyketyl sodium phosphate, isopropyl hydroxyketyl ether, glycol stearate, triisononanoïne, octyl cocoate, polyacrylamide, isoparaffin, laureth-7, a carbomer, propylene glycol, glycerol, bisabolol, a dimethicone, sodium hydroxide, PEG 30-dipolyhydroxysterate, capric / capryl triglycerides, cetearyl octanoate, dibutyl adipate, grape seed oil, jojoba oil, magnesium sulfate, EDTA, cyclomethicone, xanthan gum, citric acid, sodium lauryl sulfate, waxes and mineral oils, isostearyl isostearate, propylene glycol dipelargonate, propylene glycol isostearate, PEG 8, beeswax, hydrogenated palm heart oil glycerides, oil glycerides hydrogenated palm, lanolin oil e, sesame oil, cetyl lactate, lanolin alcohol, castor oil, titanium dioxide, lactose, sucrose, low density polyethylene, isotonic saline solution, and mixtures thereof.

Many cosmetically active ingredients are known to those skilled in the art to improve the health and / or the physical appearance of the skin (including the scalp) and / or of the mucous membranes. Those skilled in the art know how to formulate cosmetic, nutraceutical or dermatological compositions in order to obtain the best effects. On the other hand, these compounds can have a synergistic effect when combined with each other. These combinations are also covered by the present invention. The CTFA Cosmetic Ingredient Handbook, Second Edition (1992) describes various cosmetic and pharmaceutical ingredients commonly used in the cosmetic and pharmaceutical industry, which are particularly suitable for topical use. Examples of these classes of ingredients include, but are not limited to, the following compounds: abrasives, absorbents, cosmetic compounds such as perfumes, pigments, dyes, essential oils, astringents, etc. (for example: clove oil, menthol, camphor, eucalyptus oil, eugenol, menthyl lactate, witch hazel distillate), anti-acne agents, anti-flocculants, anti-foam agents, antimicrobial agents (for example: iodopropyl butylcarbamate), antioxidants, binders, biological additives, buffering agents, blowing agents, chelating agents, additives, biocidal agents, denaturants, thickeners, and vitamins, and derivatives or equivalents thereof, film-forming materials, polymers, opacifying agents, pH adjusters, reducing agents, depigmenting or brightening agents (for example: hydroquinone, kojic acid, ascorbic acid, magnesium ascorbyl phosphate, ascorbyl glucosamine), conditioning agents (eg: humectants).

The cosmetic composition may also comprise other cosmetic and / or nutraceutical agents having the same properties and inducing a synergistic effect or not with the extract according to the invention, or cosmetic agents with complementary effects. They may for example be anti-wrinkle ingredients, agents stimulating the activity or proliferation of fibroblasts, agents stimulating molecules of the extracellular matrix.

They can also be anti-aging active ingredients and / or tightening agents for a synergistic effect with the extract of Tamarindus indica. Advantageously, they are active ingredients increasing the gene and / or protein expression of collagen or active ingredients preventing the degradation of collagen such as retinol, vitamin C, an extract of Davilla rugosa marketed under the name of Collguard ™ by BASF Beauty Care Solutions, an extract of Hibiscus abelmoschus seed marketed under the name Linefactor ™, a peptide marketed under the name Dermican ™ by the applicant or the products marketed under the names of Matrixyl ™, Matrixyl 3000 ™ and Regestril ™ by the company Sederma or Neuroguard by the company Codif, or else Etemaline ™ by the company Silab.

The tensioning agents which can be used in the invention can be chosen from synthetic polymers, such as polyurethane latexes or acrylic latexes, polymers of natural origin, in particular polyholosides in the form of starch or in the form of carrageenans, alginates, agars, gellans, cellulose polymers and pectins; proteins and hydrolysates of vegetable proteins; mixed silicates; wax microparticles; colloidal particles of inorganic filler chosen for example from silica, silica-alumina composites; as well as their mixtures.

Finally, they may be cosmetic ingredients such as, for example, antimicrobial agents, anti-radical agents, soothing, calming or relaxing agents, agents acting on the microcirculation to improve the radiance of the complexion, in particular of the face, healing agents or slimming agents. Advantageously, the cosmetic and / or dermatological composition of the present invention also contains one or more tightening agents and / or one or more antimicrobial agents and / or one or more anti-free radical agents and / or one or more soothing agents and / or one or more agents. slimming agents and / or one or more active agents on the microcirculation.

Among the antimicrobial agents associated with the extract of T. indica in a preferred embodiment of the present invention, there may be mentioned 2,4,4'-trichloro-2'-hydroxy diphenyl ether (or triclosan), 3,4, 4'- trichlorobanilide, phenoxyethanol, phenoxypropanol, phenoxyisopropanol, hexamidine isethionate, metronidazole and its salts, miconazole and its salts, itraconazole, terconazole, econazole, ketoconazole, saperconazole, fluconazole, clotrimazole, butoconazole, oxiconazole, sulfaconazole, sulconazole, terbinafine, undecylenic acid and its salts, benzoyl peroxide, 3-hydroxy benzoic acid, 4-hydroxy benzoic acid, phytic acid, N-acetyl-L-cysteine, lipoic acid, azelaic acid and its salts, arachidonic acid, resorcinol, octoxyglycerin, octanoylglycine, caprylyl glycol, l 10-hydroxy-2-decanoic acid, famesol, phytosphingosines and mixtures thereof. The anti-radical agents can be vitamin C and its derivatives including ascorbyl glucoside, phenols and polyphenols, in particular tannins, ellagic acid and tannic acid; epigallocatechin and natural extracts containing it, in particular green tea extracts; anthocyanins; phenol acids, stilbenes; active scavengers of mono- or polycyclic aromatic compounds, tannins such as ellagic acid and indole derivatives and / or active scavengers of heavy metals such as EDTA, anti-radical active agents such as vitamin E and its derivatives such as tocopheryl acetate; bioflavonoids; coenzyme Q10 or ubiquinone.

As soothing agents forming part of the composition of the invention, use may be made of pentacyclic triterpenes, ursolic acid and its salts, oleanolic acid and its salts, betulinic acid and its salts, salts of salicylic acid. and in particular zinc salicylate, bisabolol, allantoin, unsaturated omega 3 oils, cortisone, hydrocortisone, indomethacin and beta methasone, anti-inflammatory active agents, and in particular those described in the application FR2847267, in particular the root extract of Pueraria lobata marketed under the name Inhipase ™ by the applicant, extracts of Theobroma cacao.

The active ingredients acting on the microcirculation, vasoprotectors or vasodilators, can be chosen from flavonoids, ruscogenins, nicotinates and essential oils.

The slimming active agents can in particular be chosen from agents which inhibit lipoprotein lipase such as those described in patent US2003086949 (Coletica) and in particular an extract of liana from Peru (Uncaria tomentosa); draining active agents, in particular hesperitin laurate (Flavagrum ™), or quercitin caprylate (Flavenger ™); phosphodiesterase enzyme inhibitors, adenylate cyclase activators, cAMP and / or active agents capable of trapping spermine and / or spermidine. Mention may be made of an extract of Coleus Forskohlii root, an extract of Cecropia obtusa, Uva lactuca, caffeine, forskolin, theophylline, theobromine and / or their derivatives, a hydrolyzed kappa carrageenan product called Slimexcess ™ marketed by the applicant and / or their mixtures.

A subject of the present invention is also a cosmetic care process comprising the topical application to at least one area of healthy skin and / or healthy mucosa of an extract of Tamarindus indica leaves according to the invention, the extract not being obtained by using butylene glycol as an extraction solvent, or of a cosmetic composition comprising it, to maintain and / or increase the expression of molecules of the extracellular matrix of healthy skin, healthy mucous membranes , and / or to maintain and / or increase the expression of collagen fibers and / or elastic fibers of healthy skin and / or healthy mucous membranes, and / or to maintain and / or increase the biomechanical properties of healthy skin and / or healthy mucous membranes, in particular firmness and / or elasticity, preferably firmness. A preferred embodiment of the invention relates to a cosmetic care process characterized in that it comprises the topical application to at least one area of healthy skin and / or healthy mucosa of a Tamarindus indica leaf extract. invention, to increase the expression of molecules of the extracellular matrix of healthy skin, healthy mucous membranes, and / or to increase the expression of collagen fibers and / or elastic fibers of healthy skin and / or healthy mucous membranes , and / or to increase the biomechanical properties of healthy skin and / or healthy mucous membranes, preferably firmness and / or elasticity and / or to increase the proliferation of fibroblasts in healthy skin and / or healthy mucous membranes, and / or to increase the differentiation of keratinocytes from healthy skin and / or healthy mucous membranes.

Advantageously, the extract of Tamarindus indica according to the invention, preferably in the form of a cosmetic composition according to the invention, is used in regular topical application and preferably at least once a day, advantageously twice a day, for at least 10 days, preferably for 20 days, and more preferably for at least 28 days.

Advantageously, the topical application of an extract of Tamarindus indica according to the invention or of a cosmetic composition comprising it, is carried out on all or part of the body and / or of the face and / or of the scalp, preferably the legs, thighs, arms, stomach, décolleté, neck, armpits, lips, more preferably all or part of the face, and preferably the cheeks, forehead, chin, lips, contour of eyes.

The cosmetic care process according to the invention for maintaining and / or increasing the expression of molecules of the extracellular matrix of healthy skin, of healthy mucous membranes, preferably for maintaining and / or increasing the expression of collagen fibers and / or elastic fibers of healthy skin and / or healthy mucous membranes, and / or to maintain and / or increase the biomechanical properties of healthy skin and / or healthy mucous membranes, in particular firmness and / or elasticity, preferably firmness advantageously comprises the following steps:

- The identification on the individual of an area of healthy skin and / or healthy mucosa, for which it is desired to maintain and / or increase the expression of molecules of the extracellular matrix of healthy skin, of healthy mucous membranes, and / or maintain and / or increase the expression of collagen fibers and / or elastic fibers of healthy skin and / or healthy mucous membranes, and / or maintain and / or increase the biomechanical properties of healthy skin and / or healthy mucous membranes, in particular firmness and / or elasticity, preferably firmness, and

- Topical application to this area of healthy skin and / or healthy mucosa, of a cosmetic composition comprising the extract of Tamarindus indica according to the invention, the extract not being obtained by using butylene glycol as an extraction solvent, in an amount effective to maintain and / or increase the expression of molecules of the extracellular matrix of healthy skin, healthy mucous membranes, and / or to maintain and / or increase l '' expression of collagen fibers and / or elastic fibers of healthy skin and / or healthy mucous membranes, and / or to maintain and / or increase the biomechanical properties of healthy skin and / or healthy mucous membranes, in particular firmness and / or elasticity, preferably firmness, namely in an extract content of between 1x10 ⁴ % and 10% (v / v) by volume, preferably between 1x10 ⁴ % and 5% (v / v) by volume, still advantageously between 1 × 10 ³ % and 3% (v / v) by volume, more preferably between 0.1% and 1% (v / v) by volume, relative to the total volume of the composition.

For the purposes of the present invention, the term “topical route” is understood to mean direct local application and / or vaporization of the extract or of the composition according to the invention on the surface of the zone of healthy skin and / or of healthy mucosa.

For the purposes of the present invention, the term “topically and / or orally acceptable” is understood to mean an ingredient suitable for application respectively topically and / or orally, non-toxic, non-irritant for the skin including the scalp and not inducing no allergic response, and which is not chemically unstable.

Advantageously, the method according to the invention relates to a cosmetic treatment method for maintaining and / or increasing the proliferation of fibroblasts of healthy skin and / or healthy mucous membranes, and / or for maintaining and / or increasing the differentiation of the keratinocytes of healthy skin and / or healthy mucous membranes comprising the following steps:

- The identification on the individual of an area of healthy skin and / or healthy mucosa, for which it is desired to maintain and / or increase the proliferation of fibroblasts of healthy skin and / or healthy mucous membranes, and / or to maintain and / or increase the differentiation of keratinocytes from healthy skin and / or healthy mucous membranes, and

- Topical application to this area of an area of healthy skin and / or healthy mucosa, of a cosmetic composition comprising the extract of Tamarindus indica according to the invention, the extract not being obtained by using butylene glycol as extraction solvent, in an amount effective to maintain and / or increase the proliferation of fibroblasts of healthy skin and / or healthy mucous membranes, and / or to maintain and / or increase the differentiation of keratinocytes from healthy skin and / or healthy mucous membranes, namely in an extract content of between 1x10-4% and 10% (v / v) by volume, preferably between 1x10-4% and 5% (v / v) in volume, again advantageously between 1x10-3% and 3% (v / v) by volume, more preferably between 0.1% and 1% (v / v) by volume, relative to the total volume of the composition.

A subject of the invention is also the extract of T. indica according to the invention, alone or in a dermatological composition comprising it, for its use, advantageously topically, in the treatment and / or prevention of pathologies of the disease. skin and / or mucous membranes involving loss of expression of molecules of the extracellular matrix of the skin and / or mucous membranes, and / or loss of expression of collagen fibers and / or elastic fibers of the skin and / or mucous membranes, and / or a loss of the biomechanical properties of the skin and / or the mucous membranes, in particular a loss of the firmness of the skin and / or of the mucous membranes, and / or a loss of the proliferation of fibroblasts of the skin and / or mucous membranes, and / or loss of differentiation of keratinocytes from the skin and / or mucous membranes, the extract not being obtained using butylene glycol as extraction solvent.

For the purposes of the present invention, from a dermatological point of view, the expression “treatment and / or prevention of pathologies of the skin and / or of the mucous membranes involving a loss of firmness of the skin and / or of the mucous membranes” is understood to mean a increase for therapeutic purposes in the firmness of the skin and / or of the mucous membranes which have lost firmness under the effect of pathologies of the skin and / or of the mucous membranes such as, for example, rosacea, telangiectasias.

For the purposes of the present invention, from a dermatological point of view, the term “treatment and / or prevention of pathologies of the skin and / or of the mucous membranes involving a loss of elasticity of the skin and / or of the mucous membranes” is understood to mean a increase for therapeutic purposes in the elasticity of the skin and / or mucous membranes which have lost their elasticity under the effect of pathologies of the skin and / or mucous membranes such as for example solar elastosis and / or cutis pathology laxa.

The extract according to the invention can be found in the form of a pharmaceutical composition, preferably dermatological, comprising at least one pharmaceutically and / or dermatologically acceptable excipient. In one embodiment of the invention, said composition is applied topically and / or orally, preferably topically.

Advantageously, it is present in the pharmaceutical composition, preferably dermatological, at a concentration of 1x10 ⁴ % to 10% by weight, preferably between 1x10 ⁴ % and 5% by weight, again advantageously between 1x10 ³ % and 0.5% by weight relative to the total weight of the composition, said composition further comprising at least one pharmaceutically acceptable excipient.

The invention will be better understood on reading the description of the figures and the examples which follow. Examples referring to the description of the invention are presented below. These examples are given by way of illustration and in no way limit the scope of the invention. Each of the examples is general.

The examples form an integral part of the present invention and any feature which appears new with respect to any state of the prior art from the description taken as a whole, including the examples, forms an integral part of the invention.

On the other hand, in the examples, all the percentages are given by weight, unless otherwise indicated, the temperature is expressed in degrees Celsius unless otherwise indicated, and the pressure is atmospheric pressure, unless otherwise indicated.

Examples

Example 1: Preparations of different extracts of Tamarindus indica

Example la): A quantity of 10% (w / w) by weight of dried and crushed Tamarindus indica leaves (origin Burkina Faso) relative to the total mixture of solvent and leaves was extracted in water as the sole solvent for one hour with stirring at a temperature of 80 ° C. The crude extract was centrifuged, filtered (0.45 µm). The extract is therefore obtained in liquid form.

Example 1b): A quantity of 10% (w / w) by weight of dried and crushed Tamarindus indica leaves (origin Burkina Faso) relative to the total mixture of water and leaves was macerated in water as the sole solvent for 1 hour at a temperature of 80 ° C. The crude extract was centrifuged, filtered (0.45 μm) then formulated with glycerin, for a final quantity of glycerin of 80% (v / v) by volume relative to the total volume of the extract and of the glycerin . The extract is therefore obtained in liquid form.

Example le): A quantity of 10% (w / w) weight of dried and crushed Tamarindus indica leaves (origin Burkina Faso) relative to the total water and leaves mixture was macerated in water as the sole solvent for 16 hour at a temperature of 4 ° C. The crude extract was centrifuged, filtered (0.45 mih) then formulated with glycerin, for a final quantity of glycerin of 80% (v / v) by volume relative to the total volume of the extract and of the glycerin . The extract is therefore obtained in liquid form.

Example 1d): A quantity of 10% (w / w) by weight of dried and ground Tamarindus indica leaves (origin Burkina Faso) relative to the total mixture of water and leaves is extracted with stirring and under reflux in a solvent consisting of an ethanol: water mixture (70:30; v / v) at a temperature of 80 ° C for a period of one hour. The suspension was then filtered and then the hydroalcoholic extract recovered. The alcoholic phase was evaporated in vacuo and the resulting residual aqueous phase was centrifuged and filtered. This aqueous phase was atomized in the presence of maltodextrin, in a final quantity of maltodextrin of 80% (w / w) by weight relative to the total weight of the extract and of maltodextrin. The extract is therefore obtained in the form of a powder.

Example le): A quantity of 20% (w / w) by weight of dried and ground Tamarindus indica leaves (origin Burkina Faso) relative to the total mixture of water and leaves a is extracted with stirring in water as the sole solvent for one hour at a temperature of 80 ° C. The crude extract was centrifuged, decanted, filtered (0.45 μm) then atomized in the presence of maltodextrin, in a final amount of maltodextrin of 80% (w / w) by weight relative to the total weight of the final extract. . The extract is therefore obtained in the form of a powder. Example lf): Whole leaves of Tamarindus indica (origin Burkina Faso) dried were extracted under subcritical conditions with water as the sole solvent in an extraction column with a flow rate of 5 ml / min and a solvent / leaves ratio. Tamarindus indica of 25 ml / g, at a temperature of 120 ° C, and a pressure of 16 bars. The recovered extract was dried and crushed. The extract is therefore obtained in the form of a powder.

Example 1g): Whole leaves of Tamarindus indica (origin Burkina Faso) dried were extracted under subcritical conditions with water as the sole solvent in an extraction column with a flow rate of 5 ml / min and a solvent / leaves ratio. Tamarindus indica of 25 ml / g, at a temperature of 120 ° C, and a pressure of 8 bars. The recovered extract was dried and crushed. The extract is therefore obtained in the form of a powder.

Example 2 Demonstration of the effect of an extract of Tamarindus indica according to the invention on the proliferation of fibroblasts

Protocol:

Normal human fibroblasts, that is to say non-pathological, obtained from abdominal biopsies of a healthy donor were inoculated in a 96-well plate in a defined medium (FGM) in the presence of the Tamarindus indica extract prepared according to example la) of the present invention, at a final concentration of 1% (w / w) by weight of the extract relative to the weight of the culture medium and of the extract. The same culture medium without addition of extract according to the invention was used as a control (untreated control). After 4 days of culture at 37 ° C, the culture medium was removed and discarded. A solution of 3- (4,5-dimethyl-2-thiazolyl) -2,5-diphenyl-2H-tetrazolium (MTT) bromide is deposited on the cell mats for 3 hours. The solution is then removed and replaced with dimethyl sulfoxide for 15 minutes. The Optical densities of each condition are then measured using an EnVision multi-plate reader (Perkin Elmer).

For each condition, the results are expressed by the average of the percentages of the cell viabilities relative to the average of the viabilities measured under the control conditions (untreated control) (n = 3).

Results: [Table 1]

Conclusion: The extract of Tamarindus indica according to example la) induced a stimulation of the proliferation of human fibroblasts.

Example 3: Demonstration of the effect of an extract of Tamarindus indica according to the invention on the synthesis of involucrine.

Protocol:

Normal human keratinocytes, that is to say non-pathological, from adult donors were inoculated into a growth medium (standard medium MCDB 153, Sigma-Aldrich France supplemented with 2% fetal calf serum). After 3-4 days of incubation at 37 ° C, CO2 = 5%, the growth medium was exchanged for standard MCDB 153 medium without FCS (untreated control), and with a range of concentrations of the extract of Tamarindus indica according to example la). As a positive reference, a range of CaCl2 concentrations (0.1 - 0.3 - 1 mM) was tested on human keratinocytes to calibrate the test. After 3 days of incubation at 37 ° C, CO2 = 5%, cell viability was quantified using the MTT test, and the level of synthesized involucrin was measured by an ELISA method on cell homogenates according to the recommendations of the maker.

The results were calculated relative to a range of involucrin, then they were expressed as a percentage relative to the control cell culture medium without product (untreated control). The results are presented in the form of the mean ± standard deviation from 2 tests (from 2 different keratinocyte donors) each carried out in triplicate. The activity of the product was statistically evaluated according to the Sigmaplot ™ software process. Results:

[Table 2]

The extract of Tamarindus indica according to example la) at concentrations of 0.3% (w / w) (w / w) by weight of the liquid extract relative to the weight of the culture medium and of the extract and 1% (w / w) by weight of the liquid extract relative to the weight of the culture medium and of the extract significantly increases the synthesis of involucrin by keratinocytes by 37% and 53%, respectively. The extract of Tamarindus indica according to the invention can therefore be used to maintain and / or increase the differentiation of keratinocytes from healthy skin and / or healthy mucous membranes.

Example 4: Increase in the expression of type I and V collagen by the fibroblasts in the presence of various extracts of T. indica according to the invention.

Protocol: So-called "normal" human fibroblasts, that is to say not showing any pathology, from a healthy 42-year-old donor were cultured in a defined medium (FGM) for a period of 48 hours in the presence of of the extract of Tamarindus indica according to example la) at a final concentration of 0.3% (w / w) by weight of the extract relative to the weight of the culture medium and of the extract or at 1% (w / w) by weight of the extract relative to the weight of the culture medium and of the extract, then the cell medium was removed. The same culture medium without addition of extract according to the invention was used as a control (Control). The resulting cell layer was lysed with ammonium hydroxide solution.

The assay of mature type I and V collagens was evaluated using time-resolved fluorescence intensity technology, DELF1A®, developed and patented by the applicant.

A phosphate buffered saline / bovine serum albumin saturation solution (Perkin Elmer, Courtaboeuf, France) was added to the matrix deposited before reaction with either the primary anti-collagen 1 antibody (fnterchim / Acris, Montluçon, France, LH0284 / RI 038) or the primary anti-collagen V antibody (fnterchim / Acris, Montluçon, France, LH4523 / RI 042). After rinsing in phosphate buffered saline, the secondary antibody conjugated to Europium (Perkin Elmer) was added. Finally, a revealing solution (Perkin Elmer) was added. Fluorescence intensity was read (/.cxc. 340nm / /.cm. 615nm) using an EnVision® multiple label plate reader (Perkin Elmer).

Fluorescence results were normalized to fluorescence obtained with the same cell medium in the absence of T. indica extract (Control) and were related to the amount of DNA obtained under each condition. The results presented correspond to the average of 6 trials (n = 6). (AND: Standard deviation).

Results: [Table 3]

Conclusion: The extract of Tamarindus indica according to example la) induced an increase in the protein expression of type I and V collagen in human fibroblasts cultured in vitro. The extract according to the invention can therefore be used to increase the firmness of the skin and / or of the mucous membranes.

Example 5: Increase in the expression of type IV collagen by keratinocytes in the presence of various extracts of T. indica according to the invention.

Protocol: Healthy human keratinocytes obtained from a human skin biopsy of a 50-year-old woman were seeded and cultured until confluence in a defined medium (KSFM, Life Technologies, France) at 37 ° C with 5% of C02 under 95% relative humidity.

The keratinocytes were incubated for 48 hours at 37 ° C, with 5% CO2 with or without Tamarindus indica extract according to example la) at 0.1%. The untreated control corresponds to the conditions without any product added to the culture medium and TGF b at 5 ng / mL was used as a positive reference.

The culture medium was removed and the keratinocytes underwent lysis with an ammonium hydroxide solution (Sigma, France). The wells were then saturated for 30 minutes with a solution of bovine serum albumin (BSA; Sigma France), then incubated for one hour with an anti-collagen IV monoclonal antibody diluted to 1/2500 followed by washing and incubation. an additional hour at room temperature with a secondary antibody diluted to 1/25000. The cell culture medium was then removed and replaced with a revealing fluorescent solution. The fluorescence was finally recorded at 340exc / 615em nm. The concentration of collagen IV was deduced from a standard curve of commercially available collagen solution IV. Readings were normalized to the amount of cellular DNA determined using a Picogreen kit.

The results of the synthesis of collagen IV by the keratinocytes are normalized relative to the untreated control and are expressed as an average percentage (%) ± SD. Statistical analysis was performed using the One way ANOVA Dunnet method following the recommendation of Sigmaplot software (Systat Software Inc, USA). The significance level was set at 5% (p <0.05).

Results [Table 4]

Conclusion: The extract of Tamarindus indica according to example la) induced an increase in the protein expression of type IV collagen in human keratinocytes cultured in vitro. The extract according to the invention can therefore be used to increase the firmness of the skin and / or of the mucous membranes.

Example 6: Demonstration of the effect of an extract of Tamarindus indica according to the invention on the gene expression of molecules of the extracellular matrix

Protocol:

Four different strains of primary human fibroblasts were obtained from surgical skin biopsies of female donors (between 52 and 64 years of age). The 4 or 5 passage fibroblasts were cultured in Eagle's minimum essential medium with fetal calf serum (FCS) and incubated at 37 ° C with 5% CO2.

After 24 hours of growth, the cells were treated with the extract of Tamarindus indica according to example la) at a final concentration of 0.3% (w / w) by weight of the extract relative to the weight of the medium. of culture and the extract or at 1% (w / w) by weight of the extract relative to the weight of the culture medium and of extract, and were incubated for 24 or 48 hours at 37 ° C with 5% CO 2. The control conditions correspond to the culture of fibroblasts without adding product to the basal culture medium. The absence of treatment-induced cytotoxicity was verified using the trypan blue method.

Total RNAs from fibroblast cultures were extracted with the NucleoSpin miRNA kit (Machery-Nagel), according to the instructions in the manual. The quantification and the quality control of the total RNAs were carried out by measuring the optical densities at 260 and 280 nm.

A first analysis was carried out on a mixture of the same quantity of total RNA extracted from 3 cultures of treated fibroblasts. A personalized PCR matrix dedicated to 88 strategic genes involved in the quality of the dermal extracellular matrix (Prime PCR Custom Plate 384 Wells-96 genes, Biorad) was created. A panel of genes whose expression was modulated by treatments (the extract of Tamarindus indica according to example la) was selected. To validate the genetic modulations identified, specific qRT-PCR analyzes dedicated to the selected target genes were carried out on each of the 4 fibroblast cultures. Reverse transcriptions were carried out on 0.1 μg of RNA in 20 mΐ of final mixture using SuperScript IV V1LO Master Mix (fnvitrogen). Quantitative PCRs of each gene were carried out on 2 mΐ of reverse transcription mixture diluted to 1/2 in 10 mΐ of final reaction mixture using the LC480 SYBR Green 1 Master system (Roche) and dedicated primers in an LC480 ™ thermal cycler. (Rock). Analysis of domestic genes (ACTB: actin B; SDHA: succinate dehydrogenase) was performed in parallel with the target genes. Each qPCR analysis was performed on duplicate and the average Ct values of each analysis were used for the calculation of the modulation of gene expression by the AACt method.

The results were expressed as a percentage +/- standard deviation of target gene expression in the condition treated with the extract of Tamarindus indica according to example 1a) relative to the untreated condition (at 100%) for each culture. (n = 4). Statistical analysis was performed using Mann & Whitney test against untreated condition.

Results: [Table 5]

Conclusion: The extract of Tamarindus indica according to example la) induced an increase in the gene expression of genes encoding collagen molecules and / or elastic fibers. The extract according to the invention can therefore be used to increase the biomechanical properties of healthy skin and / or healthy mucous membranes, in particular the firmness and / or elasticity and / or density of healthy skin and / or mucous membranes. healthy.

Example 7: Example of a cosmetic ingredient comprising an extract of Tamarindus indica according to the invention

[Table 6]

Example 8 Example of a cosmetic composition comprising an extract of Tamarindus indica according to the invention

The procedure is carried out according to methods known to those skilled in the art to mix together the various phases A, B, C, D below to prepare a composition according to the present invention. The proportions are expressed in% and the names in capital letters correspond to the INCI names of the ingredients.

Phase A

GLYCERYL STEARATE AND PEG- 100 STEARATE 4.00

PENTAERYTHRITYL DISTEARATE 1.50

CETEARYL ISONONANOATE 3.00

PROPYLHEPTYL CAPRYLATE 5.00

COCONUT CAPRYLATE 2.00

DICAPRYLYL CARBONATE 3.00

DIMETHICONE 1.00

Phase B

Water 64.43

Propylene glycol, phenoxyethanol, chlorphenesin, methylparaben 1, 00

Glycerin 1.57

Xanthan gum 0.20

Butylene glycol 2.00

Sodium hydroxide, water 0.15

Phase C

Carbomer 0.15

Water 10

Phase D

Tamarindus indica extract obtained according to example 1a) 1.00

Example 9: Example of a dermatological composition Example 9a): Ointment containing the extract according to the invention.

The composition below is prepared according to methods known to those skilled in the art, in particular as regards the different phases to be mixed together. The amounts indicated are in percentage by weight relative to the total weight of the composition. Ingredient * 3.00

Excipient:

Low density polyethylene 5.50 Liquid paraffin Qsp 100

* The ingredient is prepared according to example la) above. The ingredient according to the invention is sterilized and then dried before being incorporated into the composition in the form of an ointment.

Example 9b) Cream containing the extract according to example la).

[Table 7]

Phases A and B are heated to 75 ° then mixed with stirring. Once the mixture has returned to ambient temperature, phases C and D are added with stirring.

Claims

1. Cosmetic and / or nutraceutical use of an extract of Tamarindus indica leaves to maintain and / or increase the expression of molecules of the extracellular matrix of healthy skin and / or healthy mucous membranes, the extract not being obtained using butylene glycol as extraction solvent.

2. Use according to claim 1, wherein the molecules of the extracellular matrix of healthy skin and / or healthy mucous membranes are chosen from collagen fibers and / or elastic fibers of the extracellular matrix of healthy skin and / or healthy mucous membranes.

3. Use according to claim 1 or 2, for maintaining and / or increasing the expression of collagen fibers of type I, III, IV, V, VI, VII, XII, XIII, XIV, XVI, XVII, XXIV and / or XXIX, preferably of type I, III, IV, V and / or XIV, preferably type I, IV and / or V collagen fibers, more preferably type I collagen fibers.

4. Use according to any one of claims 1 to 3, in which the expression is gene expression, preferably genes chosen from LAMB1, LAMG1, NID-1, COL7A1, BGN, SPARC, COPIAI, COL3A1, COPIA, COPIAI, COU Al, COIAA1, COP, A2, COP5A1, COP5A2, COP14A1, P4HA1, P4HA2, P4HA3, P4HB.

5. Use according to any one of claims 1 to 4, for maintaining and / or increasing the biomechanical properties of healthy skin and / or healthy mucous membranes, preferably firmness and / or elasticity and / or density of the skin. healthy skin and / or healthy mucous membranes, more preferably the firmness and / or elasticity of healthy skin and / or healthy mucous membranes.

6. Use according to any one of claims 1 to 5, for maintaining and / or increasing the proliferation of fibroblasts of healthy skin and / or healthy mucous membranes, and / or for maintaining and / or increasing the differentiation of keratinocytes from the skin. healthy skin and / or healthy mucous membranes.

7. Use according to any one of claims 1 to 67, in which the extract is obtained by extraction in a polar protic solvent, preferably obtained by aqueous extraction, more preferably obtained in water as the sole solvent.

8. Use according to one of the preceding claims, wherein the extract is applied topically to healthy skin and / or healthy mucous membranes.

9. Use according to any one of the preceding claims, such that the extract is present in the cosmetic composition at a content of between lxl0 ⁴ % and 10% (v / v) by volume, preferably between lxl0 ⁴ % and 5%. (v / v) by volume, again advantageously between 1 × 10 ³ % and 3% (v / v) by volume, more preferably between 0.1% and 1% (v / v) by volume, relative to the total volume of the composition, said composition further comprising at least one cosmetically acceptable excipient.

10. Cosmetic care process characterized in that it comprises the topical application to at least one area of healthy skin and / or healthy mucosa of an extract of Tamarindus indica according to one of claims 1 or 7, or a cosmetic composition according to claim 9, to maintain and / or increase the expression of molecules of the extracellular matrix of healthy skin, healthy mucous membranes, and / or to maintain and / or increase the expression of fibers of collagen and / or elastic fibers of healthy skin and / or healthy mucous membranes, and / or to maintain and / or increase the biomechanical properties of healthy skin and / or healthy mucous membranes, preferably firmness and / or elasticity .

11. Cosmetic care process according to claim 10, characterized in that it comprises the topical application to at least one area of healthy skin and / or healthy mucosa of an extract of Tamarindus indica according to one of claim 1 or 7, or a cosmetic composition according to claim 9, for maintaining and / or increasing the proliferation of fibroblasts of healthy skin and / or healthy mucous membranes, and / or for maintaining and / or increasing the differentiation of keratinocytes healthy skin and / or healthy mucous membranes.

12. A cosmetic care process according to any one of claims 10 or 11, in which the topical application of an extract of Tamarindus indica according to one of claims 1 or 7, or of a cosmetic composition according to claim. 9, is performed on all or part of the body and / or face and / or scalp, preferably the legs, thighs, arms, stomach, décolleté, neck, armpits, lips, more preferably all or part of the face, and preferably the cheeks, the forehead, the chin, the lips, the eye area.

13. Cosmetic care process according to any one of claims 10 to 12, in which the cosmetic composition comprises the extract of Tamarindus indica according to the invention at a concentration of Ixl0 ⁴ % to 10% (v / v) by volume, preferably from 1x10 ⁴ % to 5% (v / v) by volume, again advantageously from 1x10 ³ % and 3% (v / v) by volume, again preferably between 0.1% and 1% (v / v) by volume, relative to the total volume of the composition, said composition further comprising at least one cosmetically acceptable excipient.

14. Tamarindus indica extract obtained according to one of claims 1 or 7, or of a dermatological composition comprising it, for its use in the treatment and / or prevention of pathologies of the skin and / or mucous membranes involving loss. expression of molecules of the extracellular matrix of the skin and / or mucous membranes, and / or loss of expression of collagen fibers and / or elastic fibers of the skin and / or mucous membranes, and / or loss of biomechanical properties of the skin and / or mucous membranes, in particular loss of firmness of the skin and / or mucous membranes, and / or loss of proliferation of fibroblasts of the skin and / or mucous membranes, and / or or loss of differentiation of keratinocytes from the skin and / or mucous membranes.

15. Tamarindus indica extract for use according to claim 14, characterized in that it is in the form of a dermatological composition comprising at least one dermatologically acceptable excipient.

16. Tamarindus indica extract for its use according to any one of claims 14 to 15, characterized in that it is present in the pharmaceutical composition, preferably dermatological, at a concentration of 1x10 ⁴ % to 10% (v / v ) by volume, preferably from 1x10 ⁴ % to 5% (v / v) by volume, again advantageously from 1x10 ³ % and 0.5% (v / v) by volume, more preferably between 0.1% and 1% ( v / v) by volume, relative to the total volume of the composition, said composition further comprising at least one pharmaceutically acceptable excipient.