FR3106756A1 - Cosmetic, nutraceutical or dermatological use of an extract of Tamarindus indicaL. and / or a composition comprising it - Google Patents

Abstract

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

Description

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

The present invention relates to the cosmetic, nutraceutical and/or pharmaceutical, in particular dermatological, use of an extract of Tamarindus indica L. and/or of a composition comprising it for maintaining and/or increasing the expression of the molecules of the matrix extracellular skin and / or mucous membranes, the extract not being obtained 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, in particular 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 a combination 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 that form a matrix-filling gel. 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 physico-chemical properties (insolubility, elasticity) after its intra- and intermolecular cross-linking by a lysyl oxidase (LOX) and its deposition on the microfibrils. Elastic fibers are responsible for the elastic property of the organs that contain them (vessels, pulmonary parenchyma, elastic cartilage, 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, the functional elastic fibers are formed by the fibroblasts of the dermis, but certain components necessary for the development of the elastic fibers have also been found in the cells of the epidermis.

The turnover rate of elastic fibers is very low in adult life, although the overall level of elastin in the skin may increase. In the newborn, the microfibrils are not all completely covered with elastin, and become so around puberty. From the age of 40, inclusions appear on the fibers, more frequent in women, then the fragmentation of the elastic fibers and their disappearance under the dermo-epidermal junction (DEJ). This splitting and/or this disappearance under the JDE results in the loss of elasticity of the skin and the formation of wrinkles. The synthesis of non-functional elastic fibers is observed during photoaging, but this increase is accompanied by the accelerated loss of elastic fibers under the JDE.

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 main collagen in the skin.

Fibrillary collagen is formed from procollagen fibers, then assembled into a network, then stabilized by cross-linking. It is collagen that gives tissues their mechanical resistance, and consequently helps to maintain their firmness.

As the skin ages, the level of collagen decreases overall, which leads to a loss of firmness. This phenomenon is all the more pronounced as 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 extra-cellular matrix, contributes to a reduction 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 indica L. commonly called tamarind or tamarind is a 12-15 m tree that can reach 30 m in height, distributed nowadays in a large number of warm regions of the globe. It most likely originated in tropical Africa (particularly in Burkina Faso) from where it spread in the past to all the warm 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 immature leaves, flowers and pods are also edible. The leaves and flowers are used in the preparation of curries, salads, stews and soups in many countries, especially in times of food shortage but also as a mordant in dyeing.

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

Various studies carried out on the extracts of leaves of T. indica have highlighted the antioxidant properties of methanolic extracts of leaves, the antiglycation properties of ethanolic extracts of a residue obtained after extraction by ethyl acetate of leaves of Tamarindus indica , the anti-bacterial and antifungal activities of the aqueous, acetone and ethanolic extracts of stem bark and leaves of Tamarindus indica of Niger origin and the anti-inflammatory and antinociceptive activities of a hydro-ethanolic extract (ethanol 95%- water 5%) of leaves of Tamarindus indica of Indian origin.

Finally, patent application US2012189725 discloses the inhibitory effect of elastase, TNF-α and MMP1 activities of a Tamarindus indica leaf extract obtained with butylene glycol as extraction solvent.

Thus, to the applicant's knowledge, no prior art discloses or suggests the cosmetic, nutraceutical and/or pharmaceutical, in particular dermatological, use of an extract of Tamarindus indica and/or of a composition comprising it to maintain and/ or increase expression of extracellular matrix molecules of skin, mucous membranes, and/or to maintain and/or increase 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 mucous membranes, and/or to maintain and/or increase fibroblast proliferation of skin and/or mucous membranes, and/or to maintain and / or increase the differentiation of keratinocytes of the skin and / or mucous membranes, the extract not being obtained by 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 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 an 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 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, preferentially firmness and/or elasticity.

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

DETAILED DESCRIPTION

Within the meaning of the present invention, the term “cosmetic use and/or composition” means a non-pharmaceutical use and/or composition, that is to say which is not intended for therapeutic use and is applied to a part of the body said to be healthy, in particular on an area of the skin and/or mucous membranes said to be healthy.

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.

Within the meaning of the present invention, the term "healthy skin", "healthy mucosa" means a zone of skin, of mucosa on which the extract according to the invention is applied and called "non-pathological" by a dermatologist, that is to mean having no infection, scarring, disease or skin condition such as candidiasis, impetigo, psoriasis, eczema, acne or dermatitis, or sores or injuries.

According to the invention, the term “mucosa(s)” denotes the ocular mucosa, the vaginal mucosa, the uro-genital mucosa and/or the buccal mucosa, in particular buccal, labial and/or gingival, preferentially, the ocular mucosa and / or mouth, and still preferentially, the labial and / or ocular mucosa.

Within the meaning of the present invention, the term "maintaining and / or increasing the expression of molecules of the extracellular matrix", 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, in relation 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 extracellular matrix molecules.

Within the meaning 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, these are collagen fibers and/or elastic fibers of the extracellular matrix of healthy skin and/or healthy mucous membranes.

Thus, the present invention aims more particularly at maintaining and/or increasing the expression of collagen fibers and/or elastic fibers of the extracellular matrix of healthy skin and/or healthy mucous membranes.

Within the meaning of the present invention, the term "maintain and / or increase the expression of collagen fibers and / or elastic fibers of healthy skin and / or healthy mucous membranes" prevent a decrease and / or increase the level of gene expression, i.e. 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 the 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 by at least 40%, preferentially by at least 200% and even more preferentially by at least 400%, relative to the level of gene and/or protein expression of the collagen fibers and/or of the elastic fibers measured in the absence of the Tamarindus indica extract 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 by at least 20%, preferably by at least 40% and even more preferably by at least 50%, relative to the level of gene and/or protein expression of the collagen fibers and/or the elastic fibers measured in the absence of the Tamarindus indica extract according to the invention.

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

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

Within the meaning of the present invention, the term “type I collagen protein” means the collagen protein present in the skin, the cornea, the tendons, the ligaments, the bones, more preferably still in the skin. Thus, in a preferred embodiment of the invention, the extract of the Tamarindus indica plant 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 mode, it is the increase in the gene expression of the collagen fibers of one or more genes chosen from LAMB1 (coding for the Laminin subunit β 1 protein), LAMG1 (coding for the Laminin subunit β 1 protein γ 1), NID-1 (coding for Nidogen 1 protein), COL7A1 (coding for Collagen Type VII α 1 Chain protein), BGN (coding for Biglycan protein), SPARC (coding for Secreted protein acidic & cysteine protein rich), COL1A2 (coding for the Col I α 2 chain protein), COL3A1 (coding for the Col III α 1 chain protein), COL1A (coding for the Col I α 1 chain protein), COL1A2 (coding for the Col I α 2 chain), COL3A1 (coding for Col III α 1 chain protein), COL4A1 (coding for Collagen Type IV α 1 chain protein), COL4A2 (coding for Collagen Type IV α 2 chain protein), COL5A1 (coding for the Col V α 1 chain protein), COL5A2 (coding for the Col V α 2 chain protein), COL14A1 (coding for the Col XIV α 1 chain protein), P4HA1 (coding for the Prolyl 4-Hydroxylase Subunit α 1 protein), P4HA2 (coding for Prolyl 4-Hydroxylase Subunite α 2 protein), P4HA3 (coding for Prolyl 4-Hydroxylase Subunite α 3 protein), P4HB (coding for Prolyl 4-Hydroxylase Subunite β protein ) .

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

In a preferential mode of the invention, it is an increase in the level of protein expression of type I collagen of at least 50%, preferentially of at least 200% and even more preferentially 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 immunomarking, for example according to the method as presented in example 4 and/or 5.

Within the meaning of the present invention, the term “immunolabeling” means the technique consisting in attaching an antibody specific for a given collagen protein, preferably type I collagen protein, to said protein, with a view to revealing it by microscopic observation. Advantageously, the revelation is done by confocal microscopy.

According to another advantageous mode, 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, preferentially the firmness and/or the elasticity and/or the density of the healthy skin and/or of the healthy mucous membranes, again preferentially the firmness and/or the elasticity of the healthy skin and/or of the healthy mucous membranes.

Within the meaning of the present invention, the term “biomechanical properties of healthy skin and/or healthy mucous membranes” means the firmness and/or the elasticity and/or the density and/or the resistance to compression and/or the resistance to 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 the elasticity and / or the density of the healthy skin and / or the healthy mucous membranes, in particular the healthy dermis, more preferably it is the firmness and / or the elasticity of healthy skin and/or healthy mucous membranes, in particular 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, torsion, suction, indentation, levatometry, ballistometry, by high-resolution ultrasound scanner or elastography, preferably by indentation, by high-resolution ultrasound scanner.

Within the meaning of the present invention, the term "maintaining and / or increasing the biomechanical properties of healthy skin and / or healthy mucous membranes" means preventing a decrease and / or increasing the biomechanical properties of healthy skin and / or healthy mucous membranes , preferably the firmness and/or the elasticity of the healthy skin and/or of the healthy mucous membranes, in particular of the healthy dermis, treated with the extract according to the invention with respect to the biomechanical properties of the healthy skin and/or of the 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%, of the biomechanical properties of healthy skin and/or healthy mucous membranes treated with the Tamarindus indica extract according to the invention compared to the biomechanical properties healthy skin and/or healthy mucous membranes measured in the absence of the Tamarindus indica extract according to the invention.

In particular, the maintenance and / or increase of the biomechanical properties of healthy skin makes it possible to limit and / or reduce the slackening of healthy facial skin, in particular in the cheeks and / or the angular area of the face (commonly called rounded face or V face).

Within the meaning of the present invention, from a cosmetic point of view, the term "maintaining and/or increasing the firmness of healthy skin and/or healthy mucous membranes", preventing the reduction and/or increasing for aesthetic purposes, the firmness 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. time-induced aging, which is found for example in so-called mature skin, i.e. skins of people over 40 years old. These can be cellular stress, non-pathological physiological variations such as a change in diet, hormonal variations, especially 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 attacks.

The increase in firmness can be measured using conventional methods, in particular by in vivo measurement using the fringe projection method using a Cutometer®, a DensiScore ® , a torquemeter or even a DynaSKIN combined with a DermaTOP.

Within the meaning 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", preventing the reduction and/or causing the increase for aesthetics, 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, i.e. to say chrono-induced aging, which is found for example in so-called mature skin, that is to say the skin of people over 40, cellular stress, non-pathological physiological variations such as a change diet, hormonal changes, 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 attacks. .

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

The Tamarindus indica extract according to the invention can be used 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 of the skin healthy and/or healthy mucous membranes.

Unless otherwise specified, “fibroblasts” and/or “keratinocytes” means “healthy” fibroblasts and/or “healthy” keratinocytes, i.e. not showing any pathology.

Within the meaning of the present invention, the term “increase the proliferation of fibroblasts” means an increase in the proliferation of fibroblasts of at least 5%, preferentially of at least 10% and even more preferentially of at least 20%, by relative to the level of proliferation of the fibroblasts measured in the absence of the extract of T. indica according to the invention.

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

For the purposes of the present invention, the term "increase the differentiation of keratinocytes" means an increase in the differentiation of keratinocytes by at least 5%, preferentially by at least 10% and even more preferentially by at least 20%, relative to the level of keratinocyte differentiation measured in the absence of the T. indica extract 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 involucrin and/or filaggrin. Advantageously, it is carried out by in vitro measurement, by measuring the synthesis of involucrin, for example according to the method as presented in Example 3.

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

The extract according to the invention can be any extract of all or part of the Tamarindus indica plant. The following can in particular be chosen: the root, the bark, the fruit, the seed, the germ, the aerial parts, in particular the flower, the stem, the branches, the leaves and/or mixtures thereof. The extract according to the invention is preferentially an extract of aerial part, more preferentially of leaves of Tamarindus indica.

The extract according to the invention may be obtained by any plant extraction method known to those 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-release, extraction assisted by enzymes, extrusion, extraction using ultrasound and/or microwaves. Preferably, the extraction is carried out by maceration.

The part of the plant is preferentially 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 protic polar 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 sole solvent.

Within the meaning of the present invention, the term “protic polar solvent” means a solvent possessing 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. Within the meaning of the present invention, the term "extract obtained by aqueous extraction" means 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 containing no alcohol, preferably containing only water.

In an alternative embodiment, the extraction may be carried out in the presence of a solvent of the C ₆ -C ₁₆ dialkyl carbonate type. Mention may be made, by way of example of this embodiment, of obtaining an extract according to the invention by adding the part of Tamarindus indica to be extracted, preferably 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 may 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%, more advantageously it will be 1% by weight relative to the total weight of the extract and solvent mixture.

According to another embodiment, the extraction may 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 the leaves (plant/solvent ratio=1/10) with stirring to the extraction solvent (a mixture of water , coconut water) whose pH was adjusted to 3 or 4, for 1 hour at 80° C. in a closed environment to prevent 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 may be obtained by extraction under supercritical conditions (CO ₂ ) in the presence of a co-solvent. Advantageously, the co-solvent will be ethanol.

In another alternative embodiment of the invention, the extraction may be carried out by aqueous extraction under subcritical conditions.

By extraction in "subcritical conditions" is meant an 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 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 ambient temperature, i.e. a temperature of 20°C. In a preferential embodiment of the invention, the extraction will be carried out at a temperature of 60°C to 90°C, preferentially from 70°C to 85°C, more preferentially 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, more preferably from 4°C to 20°C, more advantageously at room temperature, that is to 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 else 120°C, 145°C then 170°C.

The extraction can be carried out for a period ranging from 1 hour to 20 hours, preferably during a period ranging from 2 hours to 16 hours. Preferably again, said extraction is carried out for a period of 1 hour.

The Tamarindus indica extract can be obtained by extraction of 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 from 5% to 10%, and even more advantageously 10%, by weight relative to the total weight of the material and of the extraction solvent by weight of dry matter of at least a part of the plant relative to the total weight of the mixture consisting solvent and plant part (w/w). Preferably, the extract is obtained by extracting an amount of 10% by weight of dry matter from at least part of the plant, relative to the total weight of the mixture consisting of the solvent, preferably water, and the plant (p/p).

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 using 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 , preferably of leaves of Tamarindus indica, 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 sole solvent of an amount of 10% (w / w) of crushed leaves relative to the total weight of the solvent and 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 1a).

- In a second embodiment of the invention, the extract prepared according to the previous procedure is diluted with glycerin, so as to obtain a final glycerin concentration of 80% (v / v) by volume of glycerin per relative to the total volume of glycerin and extract. The extract is obtained in liquid form under the conditions described in example 1b).

- In a 3 ^rd embodiment of the invention, the extract is obtained by maceration in water as sole solvent of an amount of 10% (w / w) by weight of crushed leaves relative to the total weight of the solvent and 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 to obtain a final glycerin concentration of 80% (v/v) by volume of glycerin relative to the total volume of glycerin and extract. The extract is obtained in liquid form under the conditions described in example 1c).

- In a ^4th 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 and then the hydroalcoholic extract recovered. The alcoholic phase is evaporated under vacuum and the residual aqueous phase obtained is centrifuged and filtered. This aqueous phase is 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 in solid form, under the conditions described in example 1d ).

- In a ^5th embodiment of the invention, the extract is obtained by extraction in water as sole solvent of an amount of 20% (w / w) by weight of crushed leaves relative to the weight of the solvent and 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 amount 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 example 1e).

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

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

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 sufficient pressure for the water to be in its liquid state, i.e. 18 bar. 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 ) to an application by topical route and/or orally, more preferably to an application by topical route.

When it is used alone in the form of active ingredient, the extract according to the invention is preferentially 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 mixtures thereof.

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

Advantageously, the extract according to the invention is soluble and/or dissolved 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, preferentially 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 nutraceutically acceptable excipient. Preferably, it is incorporated into a cosmetic composition comprising at least one cosmetically acceptable excipient.

Within the meaning 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 preferential 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, preferentially between 1x10 ^-4 % and 5% (v/v) by volume, again advantageously between 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.

The cosmetic and/or dermatological composition of the invention may be chosen from an aqueous or oily solution, a cream or an aqueous 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; a 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 make-up powders, sticks or sticks, in particular in the form of lipstick.

It can also be a make-up product or a make-up 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 the face and/or the scalp, preferably the legs, thighs, arms, stomach, the neckline, the neck, the armpits, the lips, again preferentially all or part of the face, and preferentially the cheeks, the forehead, the chin, the lips, the eye contour.

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 tone.

Thus, advantageously the cosmetic composition is 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 the mucous membranes.

Alternatively, the extract according to the invention may be in any galenic form conventionally used for oral application, in particular in the form of a nutraceutical active ingredient and/or 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, matting agents, stabilizers , antioxidants, texture agents, shine agents, film-forming agents, solubilizers, pigments, dyes, perfumes and sunscreens. 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 their derivatives, protein hydrolysates, Jojoba oil and its derivatives, lipo/water-soluble esters, betaines, aminoxides, plant extracts, sucrose esters, titanium dioxides, glycines, and parabens, and again preferably from the group consisting of butylene glycol, steareth-2, steareth-21, stearyl ether glycol-15, cetearyl alcohol, phenoxyethanol, methylparaben, ethylparaben, propylparaben, butylparaben, butylene glycol, natural tocopherols, glycerin, dihydroxycetyl sodium phosphate, isopropyl hydroxycetyl ether, glycol stearate, triisononanoin, octyl cocoate, polyacrylamide, isoparaffin, laureth-7, a carbomer, propylene glycol, glycerol, bisabolol, a dimethicone, sodium hydroxide, PEG 30-dipolyhydroxysterate, capric/caprylic triglycerides, cetearyl octanoate, dibutyl adipate, grapeseed oil, jojoba oil, magnesium sulfate, EDTA, cyclomethicone, xanthan gum, citric acid, sodium lauryl sulphate, waxes and mineral oils, isostearyl isostearate, propylene glycol dipelargonate, propylene glycol isostearate, PEG 8, beeswax , hydrogenated palm heart oil glycerides, hydrogenated palm oil glycerides, lanolin oil, sesame oil, cetyl lactate, lanolin alcohol, castor oil, titanium, 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 physical appearance of the skin (including the scalp) and/or mucous membranes. A person skilled in the art knows how to formulate cosmetic, nutraceutical or dermatological compositions 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, compounds for aesthetic purposes such as perfumes, pigments, dyes, essential oils, astringents, etc. (for example: clove oil, menthol, camphor, eucalyptus oil, eugenol, menthyl lactate, distillate of witch hazel), anti-acne agents, anti-flocculating agents, anti-foaming agents, antimicrobial agents (for example: iodopropyl butylcarbamate), antioxidants, binders, biological additives, buffering agents, swelling 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 lightening 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.

These may be, for example, anti-wrinkle ingredients, agents stimulating the activity or proliferation of fibroblasts, agents stimulating the molecules of the extracellular matrix.

They may also be anti-ageing active ingredients and/or tensor agents for a synergistic effect with the Tamarindus indica extract. Advantageously, these 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 Collguard ^TM by BASF Beauty Care Solutions, a Hibiscus abelmoschus seed extract marketed under the name Linefactor ^TM , a peptide marketed under the name Dermican ^TM by the plaintiff or the products marketed under the names Matrixyl ^TM , Matrixyl 3000 ^TM and Regestril ^TM by the company Sederma or Neuroguard by the company Codif, or else Eternaline ^TM 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, cellulosic polymers and pectins; vegetable proteins and protein hydrolysates; mixed silicates; wax microparticles; colloidal particles of inorganic filler chosen for example from silica, silica-alumina composites; as well as mixtures thereof.

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 tensioning agents and/or one or more antimicrobial agents and/or one or more antiradical agents and/or one or more soothing agents and/or one or more slimming agents and/or one or more agents active on the microcirculation.

Among the antimicrobial agents associated with the extract of T. indica in a preferential mode of the present invention, mention may be made of 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, l phytic acid, N-acetyl-L-cysteine, lipoic acid, azelaic acid and its salts, arachidonic acid, resorcinol, octoxyglycerin, octanoylglycine, caprylyl glycol, 10-hydroxy-2-decanoic acid, farnesol, 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 agents that trap mono- or polycyclic aromatic compounds, tannins such as ellagic acid and indole derivatives and/or active agents that trap 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, omega-3 unsaturated 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, the extracts of Theobroma cacao.

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

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

The present invention also relates to a cosmetic care process comprising the topical application to at least one zone of healthy skin and/or healthy mucous membrane of an extract of Tamarindus indica according to the invention, the extract not not being obtained by using butylene glycol as an extraction solvent, or 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, preferentially firmness.

Advantageously, the Tamarindus indica extract 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 even 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 the face and/or the scalp, preferentially the legs, the thighs, the arms, the stomach, the décolleté, the neck, the armpits, the lips, still preferentially all or part of the face, and preferentially the cheeks, the forehead, the chin, the lips, the outline of the 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, healthy mucous membranes, preferentially 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, preferentially firmness advantageously comprises the following steps:

- The identification on the individual of an area of healthy skin and / or healthy mucous membrane, for which one wishes to maintain and / or increase the expression of molecules of the extracellular matrix of healthy skin, 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, including firmness and/or elasticity, preferentially firmness, and

- The topical application to this area of healthy skin and/or healthy mucous membrane, of a cosmetic composition comprising the extract of Tamarindus indica according to the invention, the extract not being obtained by using butylene glycol as as extraction solvent, in an amount effective to maintain and/or increase expression of extracellular matrix molecules of healthy skin, healthy mucous membranes, and/or to maintain and/or increase 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, preferentially the firmness, namely in an extract content of between 1x10 ^-4 % and 10% (v / v) by volume, preferentially between 1x10 ^-4 % and 5% (v / v) by volume, again advantageously between 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.

Within the meaning of the present invention, the term “topical route” means the direct local application and/or the vaporization of the extract or of the composition according to the invention on the surface of the zone of healthy skin and/or healthy mucosa.

Within the meaning of the present invention, the term “topically and/or orally acceptable” means an ingredient suitable for application respectively by topical and/or oral route, non-toxic, non-irritating for the skin including the scalp and not which does not induce an 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 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 mucous membrane, for which one wishes to maintain and / or increase the proliferation of fibroblasts of healthy skin and / or healthy mucous membranes, and / or to maintain and/or increase keratinocyte differentiation from healthy skin and/or healthy mucous membranes, and

- The topical application to this zone of a zone of healthy skin and/or of healthy mucous membrane, 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 proliferation of fibroblasts from 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) 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.

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 for the treatment and/or prevention of pathologies of the skin and/or mucous membranes involving loss of expression of extracellular matrix molecules 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, including loss of skin and / or mucous membrane firmness, and / or loss of skin fibroblast proliferation and/or mucous membranes, and/or loss of keratinocyte differentiation of the skin and/or mucous membranes, the extract not being obtained using butylene glycol as the extraction solvent.

Within the meaning of the present invention, from a dermatological point of view, the term “treatment and/or prevention of pathologies of the skin and/or mucous membranes involving a loss of firmness of the skin and/or mucous membranes” means a increase for therapeutic purposes of the firmness of the skin and / or mucous membranes which have lost firmness under the effect of pathologies of the skin and / or mucous membranes such as for example rosacea, telangiectasias.

Within the meaning of the present invention, from a dermatological point of view, the term “treatment and/or prevention of pathologies of the skin and/or mucous membranes involving a loss of elasticity of the skin and/or mucous membranes” means a increase for therapeutic purposes of 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 in scope.

The examples form an integral part of the present invention and any feature appearing 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 1a): An amount 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 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) weight of dried and ground 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 (0.45μm) and then formulated with glycerin, for a final amount of glycerin of 80% (v/v) by volume relative to the total volume of the extract and glycerin . The extract is therefore obtained in liquid form.

Example 1c): A quantity of 10% (w / w) 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 16 hour at a temperature of 4°C. The crude extract was centrifuged, filtered (0.45μm) and then formulated with glycerin, for a final amount of glycerin of 80% (v/v) by volume relative to the total volume of the extract and 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 under vacuum and the residual aqueous phase obtained was centrifuged and filtered. This aqueous phase was 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 maltodextrin. The extract is therefore obtained in the form of a powder.

Example 1e ) : An amount of 20% (w / w) by weight of dried and crushed Tamarindus indica leaves (origin Burkina Faso) relative to the total mixture of water and leaves a is extracted with stirring in water as a single solvent for one hour at a temperature of 80°C. The crude extract was centrifuged, decanted, filtered (0.45μm) 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 1f) : Dried whole leaves of Tamarindus indica (origin Burkina Faso) were extracted under subcritical conditions with water as sole solvent in an extraction column with a flow rate of 5 ml/min and a solvent/leaf ratio of Tamarindus indica of 25 ml / g, at a temperature of 120°C, and a pressure of 16 bars. The recovered extract was dried and ground. The extract is therefore obtained in the form of a powder.

Example 1 g ) : Dried whole leaves of Tamarindus indica (origin Burkina Faso) were extracted under subcritical conditions with water as sole solvent in an extraction column with a flow rate of 5 ml / min and a solvent / leaf ratio of Tamarindus indica of 25 ml / g, at a temperature of 120°C, and a pressure of 18 bars. The recovered extract was dried and ground. The extract is therefore obtained in the form of a powder.

Example 2: Highlighting 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 seeded in a 96-well plate in a defined medium (FGM) in the presence of the Tamarindus indica extract prepared according to Example 1a) 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 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 layers for 3 hours. The solution is then removed and replaced with dimethylsulfoxide for 15 minutes. The optical densities of each condition are then measured using an EnVision multiplate reader (Perkin Elmer).

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

Results :

Mean Standard deviation Control 100 1 Tamarindus indica leaf extract according to example 1a) at 1% (w/w) 120.06 0.14

Conclusion : The extract of Tamarindus indica according to example 1a) induced a stimulation of the proliferation of human fibroblasts.

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

Protocol:

Normal, that is to say non-pathological, human keratinocytes from adult donors were seeded in 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 with standard medium MCDB 153 without FCS (untreated control), and with a range of concentrations of the extract of Tamarindus indica according to Example 1a). As a positive reference, a range of CaCl2 concentrations (0.1 - 0.3 - 1 mM) were tested on human keratinocytes to calibrate the assay.

After 3 days of incubation at 37°C, CO2 = 5%, the cell viability was quantified using the MTT assay, and the level of involucrin synthesized was measured by an ELISA method on the 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 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 ^TM software process.

Results :

Results in % / Control Involucrin (ELISA on keratinocyte homogenate) Cup 1 Cup 2 Cup 3 Mean Standard deviation Witness 142 70 88 100 37 Extract according to Example 1a) at 0.3% 169 110 132 137 (p=0.07) 30 Extract according to Example 1a) at 1% 161 167 129 153 (p<0.01) 20

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

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

Protocol:

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

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

A saline phosphate/bovine serum albumin buffer solution (Perkin Elmer, Courtaboeuf, France) was added to the deposited matrix before reaction with either the anti-collagen I primary antibody (Interchim/Acris, Montluçon, France, LH0284/R1038 ) or the primary anti-collagen V antibody (Interchim/Acris, Montluçon, France, LH4523/R1042). After rinsing in phosphate buffered saline, the Europium-conjugated secondary antibody (Perkin Elmer) was added. Finally, a revealing solution (Perkin Elmer) was added. Fluorescence intensity was read (λexc. 340 nm / λem. 615 nm) using an EnVision® multi-label plate reader (Perkin Elmer).

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

Results :

Collagen I Collagen V Mean Standard deviation Mean Standard deviation Control 100 1 100 1 Tamarindus indica leaf extract according to example 1a) at 1% w/w 563.1 53.6 329 11.24

Conclusion : The Tamarindus indica extract according to example 1a) 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 different 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 to confluence in a defined medium (KSFM, Life Technologies, France) at 37°C with 5% CO2 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 1a) at 0.1%. The untreated control corresponds to the conditions without any product added in the culture medium and TGF β at 5ng/mL was used as a positive reference.

The culture medium was removed and the keratinocytes were subjected to 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 monoclonal antibody IV diluted to 1/2500 followed by washing and incubation. additional one hour at room temperature with a secondary antibody diluted to 1/25000. The cell culture medium was then removed and replaced with fluorescent developer solution. Fluorescence was finally recorded at 340exc/615em nm. The concentration of collagen IV was deduced from a standard curve of commercially available collagen IV solution. 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 with respect to the untreated control and are expressed as a mean percentage (%)±SD. The statistical analysis was carried out using the One way ANOVA Dunnet method following the recommendation of the Sigmaplot software (Systat Software Inc, USA). The significance level was set at 5% (p <0.05).

Results

Collagen IV Mean Standard deviation Control 100 1 Tamarindus indica leaf extract according to example 1a) at 1% w/w 268 11.26

Conclusion : The extract of Tamarindus indica according to example 1a) 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 extracellular matrix molecules

Protocol:

Four different strains of primary human fibroblasts were obtained from surgical skin biopsies of donor women (between 52 and 64 years old). Fibroblasts that had undergone 4 or 5 passages were cultured in Eagle's minimum essential medium with fetal bovine 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 1a) 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 the extract, and were incubated for 24 or 48 hours at 37°C with 5% CO2. The control conditions correspond to the culture of fibroblasts without addition of 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 manual instructions. 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 has been modulated by the treatments the extract of Tamarindus indica according to example 1a) was selected. To validate the identified genetic modulations, 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 μl of final mix using SuperScript IV VILO Master Mix (Invitrogen). Quantitative PCRs of each gene were performed on 2 µl of reverse transcription mix diluted 1/2 in 10 µl of final reaction mix using the LC480 SYBR Green I Master system (Roche) and dedicated primers in an LC480™ thermocycler (Rock). Housekeeping gene analysis (ACTB: actin B; SDHA: succinate dehydrogenase) was performed alongside target genes. Each qPCR analysis was performed on duplicates and mean Cts from each analysis were used to calculate the modulation of gene expression by the ΔΔCt 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) compared to the untreated condition (at 100%) for each culture (n=4). Statistical analysis was performed via the Mann & Whitney test relative to an untreated state.

Results :

target gene Mean Standard deviation COL4A1 + 403% 1 COL4A2 + 185% 1 LAMB1 + 47% 1 LAMG1 + 48% 0.3 NID-1 + 67% 0.3 COL7A1 + 123% 1 COL14A1 + 58% 0.3 BGN + 61% 0.3 COL1A1 + 102% 0.3 COL1A2 + 51% 0.3 COL3A1 + 123% 0.3 COL5A1 + 355% 0.3 COL5A2 + 78% 0.3 P4HA1 + 53% 0.3 P4HA2 + 160% 0.3 P4HA3 + 97% 1 P4HB + 54% 0.3 Elastin + 577% 0.3 Fibulin 1 + 172% 0.3 Fibrilin 1 + 51% 0.3 MFAP2 + 72% 1

Conclusion : The extract of Tamarindus indica according to example 1a) induced the increase in the gene expression of the genes coding for the molecules of collagen 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 the elasticity and/or the density of healthy skin and/or mucous membranes healthy.

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

Tamarindus indica extract according to example 1a) 0.2% Glycerin 80% Water QSP 100

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

The procedure is carried out according to the methods known to those skilled in the art for mixing 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.

Stage A

GLYCERYL STEARATE AND PEG-100 STEARATE 4.00

PENTAERYTHRITYL DISTEARATE 1.50

CETEARYL ISONONANOATE 3.00

PROPYLHEPTYL CAPRYLATE 5.00

COCO 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

Stage C

Carbomer 0.15

Water 10

Stage 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

Vehicle:

Low density polyethylene 5.50

Liquid paraffin Qsp 100

* The ingredient is prepared according to example 1a) 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 1a).

Phase Denomination Quantity (% by total weight) AT Water 79.60 AT PROPYLENE GLYCOL (AND) PHENOXYETHANOL (AND) CHLORPHENESIN (AND) METHYLPARABEN 2.50 AT Glycerin 2 AT SODIUM STEAROYL GLUTAMATE 0.5 B DICAPRYLYL CARBONATE 3 B COCO-CAPRYLATE / CAPRATE 3 B CAPRYLIC / CAPRIC TRIGLYCERIDE 3 B SUCROSE POLYSTEARATE (AND) CETYL PALMITATE 3 B PENTAERYTHRITYL DISTEARATE 1 B SODIUM POLYACRYLATE 0.7 VS Water 0.9 VS Tamarindus indica extract according to example 1a) 0.1 D Citric acid and water 0.7

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 (17)

Cosmetic and/or nutraceutical use of an extract of Tamarindus indica for maintaining and/or increasing 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 an extraction solvent. Use according to claim 1, in which 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 . Use according to claim 1 or 2, for maintaining and/or increasing the expression of type I, III, IV, V, VI, VII, XII, XIII, XIV, XVI, XVII, XXIV and/or XXIX collagen fibers , preferentially of type I, III, IV, V and/or XIV, preferentially collagen fibers of type I, IV and/or V, more preferentially collagen fibers of type I. 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, COL1A2, COL3A1, COL1A, COL1A2, COL3A1, COL4A1, COL4A2, COL5A1, COL5A2, COL14A1, P4HA1, P4HA2, P4HA3, P4HB . 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 the firmness and / or elasticity and / or density of healthy skin and/or healthy mucous membranes, more preferably the firmness and/or the elasticity of healthy skin and/or healthy mucous membranes. Use according to any one of claims 1 to 5, for maintaining and/or increasing the proliferation of fibroblasts in healthy skin and/or healthy mucous membranes, and/or for maintaining and/or increasing the differentiation of keratinocytes in healthy skin and/or healthy mucous membranes. Use according to any one of Claims 1 to 6, in which the extract is any extract of all or part of the Tamarindus indica plant, preferably an extract of Tamarindus indica leaves. Use according to any one of Claims 1 to 7, in which the extract is obtained by extraction in a protic polar solvent, preferably obtained by aqueous extraction, more preferably obtained in water as sole solvent. Use according to one of the preceding claims, in which the extract is applied topically to healthy skin and/or healthy mucous membranes. Use according to any one of the preceding claims, such that the extract is present in the cosmetic composition at 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 1x10 ^-3 % and 3% (v / v) by volume, still 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. Cosmetic care process characterized in that it comprises the topical application to at least one area of healthy skin and/or healthy mucous membrane of an extract of Tamarindus indica according to one of Claims 1, 7, or 8 , or of a cosmetic composition according to claim 10, for maintaining and/or increasing the expression of molecules of the extracellular matrix of healthy skin, healthy mucous membranes, and/or for maintaining and/or increasing 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, preferentially the firmness and/or the elasticity. Cosmetic care process according to Claim 11, characterized in that it comprises the topical application to at least one zone of healthy skin and/or healthy mucous membrane of an extract of Tamarindus indica according to one of Claims 1 ,7, or 8, or of a cosmetic composition according to claim 10, 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 of healthy skin and/or healthy mucous membranes. Cosmetic care process according to any one of Claims 11 or 12, in which the topical application of an extract of Tamarindus indica according to one of Claims 1, 7, or 8, or of a cosmetic composition according to claim 10, 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, even more preferably all or part of the face, and preferably the cheeks, the forehead, the chin, the lips, the eye contour. Cosmetic care process according to any one of Claims 11 to 13, in which the cosmetic composition comprises the Tamarindus indica extract according to the invention at a concentration of 1x10 ^-4 % to 10% (v/v) by volume, preferentially from 1x10 ^-4 % to 5% (v/v) by volume, still advantageously from 1x10 ^-3 % and 3% (v/v) by volume, still preferentially 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. Tamarindus indica extract obtained according to one of Claims 1, 7, or 8, or of a dermatological composition comprising it, for the treatment and / or prevention of pathologies of the skin and / or mucous membranes involving a loss of expression of extracellular matrix molecules in the skin and/or mucous membranes, and/or loss of expression of collagen fibers and/or elastic fibers in the skin and/or mucous membranes, and/or loss biomechanical properties of the skin and/or mucous membranes, including loss of skin and/or mucous membrane firmness, and/or loss of skin and/or mucous membrane fibroblast proliferation, and/or loss of keratinocyte differentiation of the skin and/or mucous membranes. Extract of Tamarindus indica for its use according to Claim 15, characterized in that it is in the form of a dermatological composition comprising at least one dermatologically acceptable excipient. Extract of Tamarindus indica for its use according to any one of Claims 15 to 16, characterized in that it is present in the pharmaceutical composition, preferentially dermatological, at a concentration of 1x10 ^-4 % to 10% by weight, preferentially of 1x10 ^-4 % to 5% by weight, again advantageously from 1x10 ^-3 % and 0.5% by weight, more preferably between 0.1% and 1% by weight, relative to the total weight of the composition, said composition comprising additionally at least one pharmaceutically acceptable excipient.