EP2170307A2

EP2170307A2 - Inhibition of age formation

Info

Publication number: EP2170307A2
Application number: EP08786012A
Authority: EP
Inventors: Nabil Abdul-Malak; Cécile ALTOBELLI; Eric Perrier
Original assignee: BASF Beauty Care Solutions France SAS
Current assignee: BASF Beauty Care Solutions France SAS
Priority date: 2007-07-09
Filing date: 2008-07-09
Publication date: 2010-04-07
Also published as: KR20100042279A; JP2010533142A; KR20150053817A; CN101686954B; WO2009007411A2; BRPI0813926A2; US20100316743A1; FR2918569A1; JP2014088381A; US20130078202A1; WO2009007411A3; CN101686954A; FR2918569B1

Abstract

The present invention relates to the use of an active substance that promotes the inhibition of AGE formation, for the preparation of a composition for preventing and/or combating the decrease in elastic and plastic properties of tissues, and in particular of the skin, for inhibiting AGE formation, or for preventing and/or combating protein glycation at the cutaneous level. The invention also relates to a method of screening for such active substances.

Description

INHIBITION OF TRAINING PES AGES

The invention relates to the use of at least one active principle for reducing the glycation of proteins that can be glycated in tissues, for example at the level of the skin or at the level of the tissue wall of a blood vessel or of an organ.

In particular, the present invention relates to at least one substance that can be used topically or orally to act on proteins that can be glycated in tissues.

STATE OF THE ART

It is known in the prior art that non-enzymatic glycation or glycosylation is a purely chemical and spontaneous reaction of covalently binding a carbohydrate to a peptide chain. Glycation is a fundamental mechanism of aging resulting from the attachment of free sugars to amino acids or proteins.

The glycated proteins, also called advanced glycation end products or AGEs (Advanced Glycation Endproducts), decrease in particular the flexibility, the elasticity, and the functionality of the skin.

The glycation process takes place in three stages:

1- Formation of a Schiff base resulting from the fixing of a reducing sugar (glucose or fructose) or an aldehyde on the amino residues of the protein, mainly lysine and the N-terminal amino fraction.

2- Molar rearrangement, called Amadori rearrangement resulting from isomerization of the Schiff base. The rate of formation of these Amadori products is proportional to the sugar concentration. 3- slow and irreversible accumulation, by rearrangements, hydrogen transfer and formation of highly reactive intermediates, glycation end products or products of

Maillard. This reaction results in the formation of AGEs, better known as Maillard products. The rate of formation of these compounds is independent of the sugar concentration of the medium but depends on the duration of the hyperglycemia and the rate of protein turnover. The first two stages (Schiff base and Amadori rearrangement) stabilize at a plateau and can be reversed depending on the blood glucose level. However, the third is irreversible and progresses regardless of blood glucose level.

Proteins of the extracellular matrix, whose lifespan in the body is very long, are affected by glycation. Glycation modifies the properties of these proteins, making them more resistant to proteolysis and preventing their renewal. Moreover, the

AGEs induce the formation of molecular bridges between collagen fibers, making it more rigid and less soluble. Finally, AGEs could have other actions by binding to specific receptors present in macrophages, endothelial and mesangial cells, inducing the secretion of pro-inflammatory cytokines or growth factors. The importance of protein glycation has been underlined by the effect of drugs that inhibit glycation, resulting in slower aging of some functions in experimental animals. During diabetes mellitus, there is also an exaggerated glycation of proteins, linked to the rise in blood sugar.

Aminoguanidine is a pharmaceutical substance derived from guanidine. It binds to the products of glycation forming a reactive compound, unfit for crosslinking. At the level of the kidney for example, she prevents the formation of AGEs in glomeruli and reduces by 90% the excretion of albumin in diabetics.

US patent application 2007/060533 describes the use of inhibitors of AGE formation for the treatment of diabetes and various diseases related to the formation of AGEs. This field is not therefore that of cosmetics where alterations of the skin can not reasonably be considered as pathologies related to the formation of AGEs. On the other hand, it should be noted that anthocyanins are glycosylated. The application US 2007/0003536 describes in particular the topical application of a grape seed extract (vitis vinifera) in combination with phospholipids as anti-free radical agents, but does not describe an action on AGEs. The mechanisms are very different. Grape seed extract is described as an anti-hydroxyl agent. Here again, there is no link with an activity on EGAs.

GOALS OF THE INVENTION

The main purpose of the invention is to solve the technical problem of providing active substances that make it possible to limit the formation of AGEs resulting from the chemical glycation of proteins.

The present invention is intended in particular to solve this technical problem in the context of the provision of cosmetic, dermatological or pharmaceutical compositions preventing or combating the reduction of the elastic and plastic properties of the tissues, for example at the level of the skin, or at the level of the tissue wall of a blood vessel or an organ, and in particular the skin, especially during tissue aging or diabetes. The present invention also aims to provide a method for screening active ingredients having the properties mentioned above.

The present invention is intended to solve the technical problem reliably, reproducibly, providing non-toxic active substances, particularly for the cosmetics industry, dermatology, dermo-pharmacy, or pharmacy preferably topically.

The invention particularly aims to provide active ingredients derived from plants, and in particular to provide active ingredients with low toxicity and dermatologically acceptable.

The present invention also aims to provide active ingredients whose preparation is inexpensive, and industrially reliable and simple.

DESCRIPTION OF THE INVENTION

Thus, the present invention describes the use of an active substance that promotes the inhibition of the formation of AGEs for the preparation of a composition for preventing and / or combating the reduction of the elastic and plastic properties of tissues, such as, for example at the level of the skin, or at the level of the tissue wall of a blood vessel or an organ, and in particular of the skin.

The present invention relates to the use of an active substance for the preparation of a composition for promoting inhibition of AGE formation.

The present invention relates to the use of an active substance for the preparation of a composition for preventing and / or controlling the glycation of proteins in tissues, such as for example cutaneous level, or at the level of the tissue wall of a blood vessel or organ.

Glycation is implicated in many age-related progressive diseases, such as vascular diseases (such as atherosclerosis), kidney disease, arthritis, diabetes complications, scarring, and so on. It is significant that diabetic complications due to glycation, can occur even at a younger age in diabetic individuals, whose average blood sugar concentration is higher than normal. In diabetic disease, in which the involvement of free radicals is widely documented, oxidative stress is directly related to hyperglycemia. Too much sugar in the blood oxidizes easily. This oxidation of sugars leads, among other things, to sugar / protein or glycation grafts. The increased glycated hemoglobin level in diabetes is the typical example.

Glycation products whose rate is proportional to the level of blood glucose over a long period of time, are partly responsible for tissue aging, whose decline in elastic and plastic properties is one of the causes.

Thus, the present invention relates to the use of an active substance for the preparation of a composition for preventing and / or controlling the glycation of proteins, especially at the cutaneous level, linked to the rise in blood sugar during diabetes mellitus. . The present invention relates to the use of an active substance for the preparation of a composition for preventing and / or controlling the formation of AGEs in glomeruli, and in particular reducing the excretion of albumin in diabetic subjects.

Advantageously, the composition is a cosmetic, dermopharmaceutical or pharmaceutical composition, preferably applicable by topically or administered orally, for example as a dietary supplement.

The present invention also relates to a cosmetic composition, applicable topically or as a dietary supplement (nutraceutical), for preventing and / or combating the reduction of the elastic and plastic properties of tissues, and in particular of the skin, as in the case of a fight against and / or prevention of aging of a tissue, by inhibiting the formation of AGEs at the level of the tissue, comprising as active substance a substance promoting the inhibition of the formation of AGEs, optionally in combination with a substance for promote the protection of skin proteins such as aminoguanidine or guanidine, or a glycation inhibiting agent by mineralization, such as EDTA derivatives, phytic acid, azelaic acid, or an enzyme inhibitor such as pentosidine , or a substance capable of decreasing the level of sugar available to participate in the glycation reaction, such as carnosine, ascorbic acid or alph a-tocopherol.

The present invention also relates to a pharmaceutical composition, for example applicable topically or administered orally, in particular for combating and / or preventing the decrease in the elastic and plastic properties of a tissue, and in particular cutaneous tissue, or of the tissue wall of a blood vessel or of an organ, by inhibiting the formation of AGEs at the level of the tissue, characterized in that it comprises as active substance a substance promoting the inhibition of the formation of AGEs, optionally in combination with a substance to promote the protection of tissue proteins, such as aminoguanidine or guanidine, or a glycation-inhibiting agent by mineralization, such as EDTA derivatives, phytic acid, azelaic acid, or an enzyme inhibitor such as pentosidine, or a substance capable of decreasing the sugar level available to participate in the glycation reaction, such as carnosine, ascorbic acid or alpha-tocopherol.

According to a first embodiment, the active substance promoting the inhibition of the formation of AGEs is preferably chosen from an extract of a plant, whole or part thereof, selected from an extract of Epimedes, an extract of crimped sorrel, an extract of Sarsaparilla, an extract of native liana (Davilla rugosa) Guarana, preferably the seeds, a Cachou extract, a Sylmarin extract, a pine extract, a Rhubarb extract from china , an extract of Hawthorn, leucocyanidines, for example extracted from grape seeds, and in particular leucocyanidines containing proanthocyanins of structure (I), an extract of Aréquier, an extract of Blueberry, an extract of Elderberry, an extract of Walnut, a Willow extract, a salad extract, an extract of Lespedeza and their mixtures.

Structure (I)

According to the plants, it is advantageous to use a part of the plant chosen from a root, a rhizome, a stem, a bark, a flower, a fruit, a seed, a germ, and a leaf, preferably at 1-10. % (w / w) in a solvent or a mixture of solvents, preferably a protic polar solvent, and preferably in water, an alcohol, a glycol, a polyol, a water / alcohol mixture, water / glycol, or water / polyol (such as water in mixture with ethanol, glycerol, butylene glycol or other glycols, such as xylitol etc.) from 100/0 to 0/100 (v / v). The extracts obtained are then preferably filtered or distilled to recover the soluble fraction which is then filtered. The active substance is advantageously the plant extract in a solvent, such as water, an alcohol, a polyol, a glycol, or a mixture thereof, preferably diluted to a concentration of between 0.01 and 10% (v / v ).

The plant extracts according to the present invention are preferably chosen from the group consisting of: the epimedium (Epimedium brevicornum), preferentially the leaves, the kinked sorrel (Rumex crispus), preferably the root, the leaves and / or the bark , Sarsaparilla (Smilax ornata) - The native liana (Davilla rugosa), especially its leaves

The indigenous liana extract (Davilla rugosa), in particular its leaves, is of greater interest for producing a cosmetic, pharmaceutical, in particular dermatological composition, in combination with a suitable cosmetic and / or pharmaceutical vehicle, especially dermatological vehicle. . Such a composition is particularly intended for topical application to prevent and / or fight against aging of the skin and its unsightly manifestations, in particular wrinkles, fine lines, furrows and improves the general condition of the skin. The native liana is therefore useful for performing a cosmetic or dermatological treatment of applying the vine extract of the natives to a part of the human body, preferably the skin to improve the appearance and / or condition. The present invention thus relates to the use of this indigenous liana extract for cosmetic applications for aesthetic purposes or for producing a pharmaceutical composition, especially dermatological, in particular to protect the skin against the harmful effects of environmental agents including pollutants and UV and / or repair. According to a second embodiment, the active substance promoting the inhibition of the formation of AGEs is preferably chosen from the following characterized molecules: anthraquinone and its derivatives, preferentially 1,4-anthraquinone and 1-amino-2 hydroxymethylanthraquinone, 4-hydroxychalcone, an OPC (procyanidolic oligomers), such as strawberry OPC, or leucocyanidines or proanthocyanins of structure (I), prunin, catechol, 4-aminophenol, sodium erythrosine, and plant extracts containing them, and their mixtures.

The molecule characterized according to the invention is preferably chosen from the group consisting of 1,4-anthraquinone, 1-amino-2-hydroxymethylanthraquinone, 4-hydroxychalcone, strawberry OPC, proanthocyanines of structure (I), prunin and mixtures thereof,

According to a particular embodiment, the active substance is proanthocyanine of structure (I), preferably those obtained from grape seeds, and in particular an extract thereof, said extract containing at least 90%, and preferably at least 95% by weight of the proanthocyanines of structure (I). The invention therefore relates to a plant extract containing at least 90%, and preferably at least 95% by weight of the proanthocyanines of structure (I), and preferably obtained from grape seeds.

According to a particular embodiment, the strawberry OPC is obtained from the strawberry rhizome (Fragaria vesca), preferably in the form of an extract containing a mixture of oligomers of anthocyanidins, catechins, epicatechins and procyanidines, preferentially as dimers to hexamers. Thus, the invention relates to the use as an active ingredient promoting the inhibition of the formation of AGEs of a substance chosen from an extract of a whole plant or a part thereof, chosen from: Guarana, Cachou , Sylmarin, Pine, Rhubarb of China,, Epimede, Hawthorn, Leucocyanidines, Arquier, Blueberry, Elder, Walnut, Willow, Kernel Pochard, Salad, Sarsaparilla, Liana of the natives and Lespedeza; and / or a compound selected from sodium erythrosine, 1,4-anthraquinone, catechol, 4-hydroxychalcone, 4-aminophenol, OPC (procyanidolic oligomers), such as strawberry OPC, or leucocyanidines or proanthocyanines of structure (I), prunin, and 1-amino-2-hydroxymethylanthraquinone, or a mixture of these substances, for the preparation of a cosmetic, nutraceutical or pharmaceutical composition.

Among all these active substances, those which inhibit at least 65% the formation of AGEs, and preferably at least 80%, are preferred, with reference to the inhibition produced by the aminoguanidine at 1.5 mM.

The preferred active ingredients in the compositions are selected from the group consisting of an epimedal extract (Epimedium brevicornum), preferably the leaves, a kernel sorrel extract (Rumex crispus), preferentially the root, the leaves and / or the bark, an extract of sarsaparilla (Smilax ornata), preferentially the roots, a vine extract of natives (Davilla rugosa), in particular leaves, 1,4-anthraquinone, 4-hydroxychalcone, strawberry OPC, prunin or a combination of them.

The active substance can be concentrated by lyophilization, atomization, etc.

Advantageously, said substance is used at a concentration preferably between 0.001 and 10%, preferably between 0.01 and 5% and more particularly at 1% for plant extracts, and preferably between 1.10 ^"7 and 1%, preferably between ^{1.10" 7} and 1.10 ^"lo / o, still preferentially ^{IOL" 5} and 1.10 ^"lo / o to the characterized molecules, by weight of the total composition, without this being limiting of the concentration to be used Thus the present invention relates to a cosmetic treatment method comprising the topical application, or the ingestion as a dietary supplement (nutraceutical), of a composition comprising as a cosmetic active ingredient at least one of the The present invention also relates to a method of treating the human body comprising the administration of a pharmaceutical composition mentioned above or cited hereinafter, preferably topically, to prevent and / or fight against the glycation of proteins in a tissue, especially when the blood sugar level rises and / or is high, as for example during diabetes. in particular a method for reducing the excretion of albumin in a diabetic subject.

The present invention also relates to a method for screening active ingredients that inhibit the formation of AGEs, comprising: a) contacting at least one type of glycated protein, a sugar and the substance to be screened for its activity with respect to the inhibition of the formation of AGEs; b) the selection of at least one active ingredient promoting the inhibition of the formation of AGEs. Advantageously, step a) comprises an incubation of at least one type of protein of the skin or of walls of blood vessels, for example a collagen, or of a bovine serum albumin with a reducing sugar (for example glucose, ribose or fructose) or an aldehyde under conditions allowing the formation of AGEs. Advantageously, the skin protein is human collagen, which makes it possible to verify that the active substances of the invention are active on a majority protein in the human skin.

Advantageously, step b) comprises the incubation of at least one type of protein of the skin or walls of blood vessels or of a bovine serum albumin with a reducing sugar (for example ribose, glucose, or fructose) or an aldehyde in the presence of at least one substance to be screened at a temperature of between 40 and 60 ^° C., preferably at approximately 50 ^° C. The selection of the active principle is carried out by comparing the results obtained in the presence of the active agent tested against a witness or control.

The compounds according to the present invention are prepared in the form of topical compositions, in particular of cosmetic, dermopharmaceutical, or pharmaceutical, dermatologically acceptable compositions. As a result, for these compositions, the excipient contains, for example, at least one compound chosen from the group consisting of preservatives, emollients, emulsifiers, surfactants, moisturizers, thickeners, conditioners, mattifying agents, stabilizers, antioxidants, texture agents, gloss agents, film formers, solubilizers, pigments, dyes, fragrances 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 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, dihydroxycetyl sodium phosphate, isopropyl hydroxycetyl ether, glycol stearate, triisononanoin, octyl cocoate, polyacrylamide, isoparaffin, laureth-7, carbomer, propylene glycol, glycerol, bisabolol, dimethicone, sodium hydroxide, PEG-30-dipolyhydroxystate, capric / caprylic triglycerides, cetearyl octanoate, dibutyl adipate, grapeseed oil, jojoba oil, magnesium sulfate, EDTA, cyclometh icon, xanthan gum, citric acid, sodium lauryl sulphate , waxes e mineral oils, isostearyl isostearate, propylene glycol dipelargonate, propylene glycol isostearate, PEG 8, Beeswax, glycerides of hydrogenated palm heart oil, glycerides of hydrogenated palm oil, lanolin oil, sesame oil, cetyl lactate, lanolin alcohol, castor oil, titanium dioxide, lactose, sucrose, low density polyethylene, saline isotonic solution.

Advantageously, the abovementioned compositions are formulated in a form chosen from the group consisting of an aqueous or oily solution, an aqueous cream or gel or an oily gel, in particular in a pot or in a tube, in particular a shower gel, a shampoo; a milk ; an emulsion, a microemulsion or a nanoemulsion, especially oil-in-water or water-in-oil or multiple or silicone; a lotion, in particular in a glass or plastic bottle or in a measuring or aerosol flask; a lightbulb ; a liquid soap; a dermatological bread; an ointment ; a mousse; an anhydrous product, preferably a liquid product, pasty or solid, for example in the form of a stick, especially in the form of lipstick.

The term "topical application" as used herein means applying or vaporizing the composition according to the present invention on the surface of the skin.

The terms "dermatologically acceptable", used herein, mean that the composition or components thereof are suitable for use in contact with human skin without toxicity, incompatibility, instability, allergic response, or their equivalents, undue. The terms "promoting inhibition of AGE formation" mean that the substance makes it possible to reduce the formation of an advanced glycation product (AGE) of a protein capable of glycation in the presence of a sugar. It is preferred that the substance promoting the inhibition of the formation of AGEs makes it possible, for example, to obtain a fluorescence of at least 65% of that obtained in the presence of aminoguanidine at 1.5 mM and in the absence of the active substance favoring inhibition of the formation of AGEs, for example under the conditions of Example 1 below.

Many cosmetically active ingredients are known to those skilled in the art for improving the health and / or physical appearance of the skin. Those skilled in the art know how to formulate the cosmetic or dermatological compositions to obtain the best effects. On the other hand, the compounds described in the present invention 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: abrasive, absorbents, compound for aesthetic purposes such as perfumes, pigments, dyes, essential oils, astringents, etc. (eg clove oil, menthol, camphor, eucalyptus oil, eugenol, menthyl lactate, hamelite distillate), anti-acne agents, anti-flocculants, anti-foam agents, antimicrobial agents (eg iodopropyl butylcarbamate), antioxidants, binders, biological additives, buffering agents, blowing agents, chelating agents, additives, biocides, denaturants, external analgesics, materials forming films, 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 for example: humectants), skin-soothing agents and / or healing agents (eg panthenol and its derivatives (eg ethyl panthenol), aloe vera, pantothenic acid and its derivatives, allantoin, bisabolol, and dipotassium glycyrrhizinate), thickeners, and vitamins, and derivatives or equivalents thereof.

It is particularly interesting to combine the active substances that inhibit glycation with an active substance that makes it possible to limit the presence of AGEs, by reversing the Maillard reaction, by promoting the deglycation of AGEs, or by promoting the reversal of the Maillard reaction. AGEs, and in particular those selected from the group consisting of: - Aye Wiwii (Maprounea guyanensis), and preferably the leaves

- Jamaica vervain (Stachytarpheta jamaicensis),

- Cercopia (Cercopia obtusa), and preferably the leaves and / or buds, - Pyrole (Chimaphila umbellata). 3,5-Dimethoxy-4-hydroxycinnamic acid (sinapic acid)

Trans-3,3 ', 4', 5,7-Pentahydroxyflavane (catechin)

- oxindole

3,4-dihydroxyphenylacetic acid (DOPAC) 1,3,5-trihydroxybenzene (phloroglucinol)

It is also particularly advantageous to combine the active substances according to the present invention with other active agents with complementary properties to further improve the efficacy against cutaneous aging, against the reduction in flexibility, and / or plasticity, and / or elasticity, and / or the functionality of the skin. Advantageously, these other assets are chosen from:

the active substances stimulating cell proliferation and / or differentiation, having an anti-aging effect, in particular the following molecules: NGF, alpha-MSH, beta-endorphin or their derivatives, in particular those described in the patent application FR2857874,

the active substances protecting fibroblast growth factor (FGF), in particular FGF2, in particular those described in the patent application on behalf of the Applicant published under No. GB244036, in particular an extract of Hibiscus Abelmoscus, the active substances stimulating the activity and / or proliferation of fibroblasts, in particular a fermented soybean peptide, in particular that marketed by the Applicant under the name Phytokine ™, advantageously in combination with an extract of Hibiscus Abelmoscus.

the active substances stimulating Hyaluronase synthase, in particular HAS2, in particular those described in patent FR2893252,

the active substances stimulating the activity and / or the synthesis of Lysyl Oxidase, in particular LOX, in particular those described in the patent application FR2855968, and preferentially the dill extract, active substances having anti-inflammatory properties, such as the one inhibiting PLA2, in particular an extract of pueraria lobata roots as described in patent FR2847267 (Inhipase®);

Other objects, features and advantages of the invention will become apparent to those skilled in the art following the reading of the explanatory description which refers to examples which are given by way of illustration only and which can not in in no way limit the scope of the invention. The examples are an integral part of the present invention and any features appearing novel from any prior art from the description taken as a whole, including the examples, form an integral part of the invention in its function and its generality. Thus, each example has a general scope.

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

EXAMPLES

Example 1: Inhibition of the formation of AGEs by plant extracts.

1- Preparation of AGEs:

The preparation of a solution containing a protein and a reducing sugar is carried out in an extemporaneous manner as follows: a solution of bovine serum albumin (BSA) at a concentration of between 1.5 .mu.M and 1.5M, preferably between 15 .mu.M and 500 .mu.m, is incubated with a reducing sugar solution such as glucose, fructose, ribose, etc., preferably with glucose, at a concentration between 0.1M and 10M and preferably between 0.5M and 5M.

2- Preparation of potentially active principles of the "plant extract" type:

The plant extracts are obtained by macerating the plants (preferably root, rhizomes, stems, bark, flowers, fruits, seeds, seeds or leaves) at 1-10% (w / w) in a solvent or a mixture of solvents, advantageously chosen from water, an alcohol, a glycol, a polyol, or a mixture of water / (alcohol, glycol or polyol) (such as ethanol, glycerol, butylene glycol and other glycols, xylitol etc.) of 100/0 at 0/100 (v / v), and preferably in water. The extracts obtained are then filtered or distilled in order to recover the soluble fraction which is then filtered at 0.45 μm preferably to obtain the extract to be tested.

3- Incubation of the AGEs with the potentially active compound and analysis of the inhibition obtained:

The protein / reducing sugar solution prepared in 1 is incubated in the presence or absence (negative control) of an extract obtained in 2, at a temperature of between 40 and 60 ^° C., preferably at about 50 ^° C. for 1 to 5 weeks. preferably for 3 weeks. The active substances obtained by extraction according to the protocol described in paragraph 2- extracts are tested at a concentration of between 0.001 and 10%, preferably between 0.01 and 5% and more particularly at

1% by weight in the protein / reducing sugar preparation. The positive control used is aminoguanidine at a concentration of between 15 .mu.M and 15OmM, preferably at a concentration of between 1.5 and 15 mM in water. The measurement of the AGEs inhibition is carried out by measuring the fluorescence (excitation wavelength between 350 and 375 nm preferably at 355 nm; emission wavelength between 420 and 450 nm, preferably at 430 nm). The inhibition is calculated by deducing the resulting value of the interaction of plant extracts with the molecules of the reaction medium (quenching).

Plant extracts that inhibit the formation of AGEs under the conditions described are:

Table 1

% inhibition = Fluorescence obtained with the active principle versus the fluorescence obtained with the positive control (aminoguanidine 1.5 mM), in%.

The leucocyanidins of the present invention are an aqueous extract (preferably by maceration in water) from grape seeds and contain 95% of proanthocyanins corresponding to the following structure (I):

Structure (I)

By way of example, mention may be made of the leucocyanidins registered under No. 84929-27-1.

Example 2 Inhibition of the Formation of AGEs by Characterized Molecules

The preparation of a protein / sugar reducing solution is carried out according to the method described in Example 1 (paragraph 1). The molecules are tested at a concentration of between 1.10 ^"and 1.10 ^{~ lo} ⁷ / o, and preferably between ^1.10" and ⁵ IOL ^"1%, and in particular ^{IOL" 1%,} for example in water or DMSO.

The molecules that inhibit the formation of AGEs under the described conditions are: Table 2

Example 3 Activity of the substances according to the present invention on the antiqlycation of human collagen

The preparation of a protein / sugar reducing solution is carried out according to the method described in Example 1 (paragraph 1), with the use of human collagen instead of BSA.

1- Preparation of human collagen

The extraction of human collagen is performed from a biopsy

Human resulting from plastic surgery. The collagen obtained in the form of a solution is incubated with a reducing sugar solution such as glucose, fructose, ribose and preferably with ribose. The incubation concentrations are the same as described in the previous examples.

20 2- Preparation and testing of potentially active principles

The potentially active principles of the "plant extract" type and of the "characterized molecule" type are used under the same conditions as those cited in Examples 1 and 2.

3- Results

The molecules that inhibit the formation of AGEs under the conditions described in a manner at least equivalent to that of the aminoguanidine used 1.5 mM are the same as those cited in Examples 1 and 2.

Table 3: Comparative Results Between BSA and Human Collagen (5% Extracts)

* see above.

Example 4 Comparison Between Proanthocvanidines According to the Present Invention and Certain Anthocvanidines of the Prior Art

In this example the inventors wanted to study the inhibition properties of the formation of AGEs of different molecules of the anthocyanin family. Table 4

% inhibition: Fluorescence obtained with the active principle versus the fluorescence obtained with the positive control aminoguanidin 1.5mM), in

%.

* see above.

Surprisingly, it has been discovered that grape seed extract containing 95% of proanthocyanidins of structure (I) according to the present invention has a very important activity compared to the glycosylated molecules of the prior art. In the prior art, different anthocyanins have been reported to be active to inhibit AGE formation. However, this finding does not generally take into account the "quenching" or noise of the signal generated by the absorption properties of certain wavelengths of the anthocyanins themselves, and is not related to an inhibitory activity of AGE formation. .

In the present invention, due consideration is given to quenching. Thus the results observed are directly attributable to the inhibition of the formation of AGEs. Example 5 Comparison with the Teaching of US Patent Application 2007/0060533, Yohikawa et al., "Novel Inhibitor of the formation of advanced αlvcation and product and aldose reducate inhibitor":

The anthocyanins of US 2007/0060533, which can be used as inhibitors can be obtained by extraction to obtain large amounts of anthocyanins including berries.

The inventors have tested the following berries in comparison with the leucosynidines of the invention:

Table 5

*see above

It is found that the leucocaynidines of the invention have an activity much greater than that of the anthocyanins which are contained in the active berries according to the prior art. Examples 6 to 11: Anti-aging composition applicable topically

In the following examples, "Products of the invention" represents the active substances according to the invention and in particular obtained according to Example 1 or 2.

Example 6 Use of the Products of the Invention in Cosmetic or Pharmaceutical Formulations of Oil-in-Water Emulsion Type

Formulation 6a:

Water qs 100

Butylene Glycol 2

Glycerin 3

Sodium Dihydroxycetyl 2

Phosphate,

Isopropyl Hydroxycetyl Ether

B Glycol Stearate SE 14 Triisononoin 5 Octyl Cocoate 6

Butylene Glycol, 2 Methylparaben, Ethylparaben, Propylparaben, pH adjusted to 5.5 D Products of the invention 0.001 - 10%

Formulation 6b:

A water qs 100

Butylene Glycol 2

Glycerin 3

Polyacrylamide, Isoparafin, 2.8

Laureth-7

B Butylene Glycol, 2

methylparaben,

Ethylparaben, Propylparaben;

2

phenoxyethanol,

methylparaben,

Propylparaben, Butylparaben,

Ethylparaben 0.5

Butylene Glycol

D Products of the invention 0.001 - 10%

Formulation 6c:

At Carbomer 0.50

Propylene Glycol 3

Glycerol 5

Water qs 100

B Octyl Cocoate 5 Bisabolol 0.30

Dimethicone 0.30

Sodium Hydroxide 1.60

Phenoxyethanol, 0.50

methylparaben,

Propylparaben, Butylparaben,

Ethylparaben

Perfume 0.30

Products of the invention 0,00

Example 7 Use of the Products of the Invention in a Water-in-Oil Formulation

A PEG 30 3 dipolyhydroxystearate

Capric Triglycerides 3

Cetearyl Octanoate 4

Dibutyl Adipate 3

Grape Seed OiI 1,5

Jojoba OiI 1,5

Phenoxyethanol, 0.5

methylparaben,

Propylparaben, Butylparaben,

Ethylparaben

B Glycerin 3

Butylene Glycol 3 Magnesium Sulphate 0.5

EDTA 0.05

Water qs 100

Cyclometh icon 1

Dimethicone 1

D Perfume 0.3

Products of the invention 0.001 - 10%

Example 8 Use of the Products of the Invention in an Aqueous Gel Formulation (Contours of the Eye, etc.)

A water qs 100

Carbomer 0.5

Butylene Glycol 15

Phenoxyethanol, Methylparaben, 0.5

Propylparaben, Butylparaben, Ethylparaben

B Products of the invention 0,001 - 10%

EXAMPLE 9 Use of the Products of the Invention in a Shampoo or Shower Gel Formulation

At Xantham Gum 0.8 Water qs 100

B Butylene Glycol, 0.5 methylparaben,

Ethylparaben, Propylparaben

Phenoxyethanol, 0.5

methylparaben,

Propylparaben, Butylparaben,

Ethylparaben

Citric acid 0.8

D Sodium Laureth Sulfate 40.0

Product of the invention 0.001 - 10%

EXAMPLE 10 Use of the Products of the Invention in a Lipid Red and Other Anhydrous Formulation

A Wax Minerals 17.0

Isostearyl Isostearate 31.5

Propylene Glycol Dipelargonate 2,6

Propylene Glycol Isostearate 1,7

PEG 8 Beewax 3.0

Hydrogenated Palm Kernel OiI 3,4 Glycerides,

Hydrogenated Palm Glycerides

Lanolin OiI 3,4

Sesame OiI 1,7

Cetyl Lactate 1.7

OiI Ore, Lanolin Alcohol 3.0 B Castor OiI qsp 100 Titanium Dioxide 3.9 CI 15850: 1 0.616 CI 45410: 1 0.256 CI 19140: 1 0.048 CI 77491 2.048

Products of the invention 0.001 - 5%

Example 11 Use of the Products of the Invention in a Tablet, Ointment or Injection Formulation

Ua formulation: tablet preparation

Excipients In grams per tablet

Lactose 0.359

Sucrose 0,240

B Active ingredient * 0.001 -0.1

* The active ingredient is obtained, for example, according to the extraction method described in the examples, followed by a drying step.

Ub formulation: preparation of an ointment

excipients

Low density polyethylene 5.5

Liquid paraffin qs 100

B Active ingredient * 0.001 -0.1

* The active ingredient is obtained, for example, according to the extraction method described in the examples, followed by a drying step. Uc formulation: preparation of an injectable formula A Excipient

Salted isotonic solution 5 ml

B Active ingredient * 0.001 -0.1 g

Phase A and Phase B are packaged in separate ampoules and mixed before use.

Claims

M

1, Use as an active ingredient promoting the inhibition of the formation of AGEs of a substance selected from the group consisting of: an extract of a plant, whole or part thereof; selected from Epimedes, Pochardseed, Sarsaparilla, indigenous vine, Guarana seeds, Cachou, Syfmarin, Pine, Rhubarb, Epimede, Hawthorn, leucocyanidines, Arecan, Blueberry, Elder, Walnut, Willow, Kernel Pochard, Salad, Sarsaparilla , and Lespedeza; and / or a compound selected from anthraquinone and its derivatives, preferably 1,4-anthraquinone and 1-amino-2-hydroxymethylanthraquinone, 4-hydroxychalcone, an OPC (procyanoid oligomers), such as strawberry OPC, leucocyanidins or proanthocyantnes of structure (I)

(I), prunin, catechol, 4-amiπophenoi, sodium erythrosine, or a mixture of these substances for the preparation of a cosmetic, nutraceutical or pharmaceutical composition.

SUBSTITUTE SHEET (RULE 26) W

2. The use as claimed in claim 1, characterized in that the composition comprises at least one active ingredient chosen from epircede extract (Epimedium brevicornum), preferentially the leaves, a kernel sorrel extract (Rumex crispus), preferentially the root, the

5 leaves and / or the bark, an extract of sarsaparilla (Smilax ornata), preferentially the roots, an extract of vine of the natives (Davilla rugosa), preferentially of leaflets, the 1,

4-anthraquinone, 4-hydroxychalcone, OPC strawberry, prunine, or a combination of them.

K)

3, Use according to any one of the preceding claims, characterized in that the composition is a cosmetic, nutraceutical or pharmaceutical composition for preventing and / or preventing reduction of the elastic and plastic properties of a fabric, such as by

For example at the level of the skin, or at the level of the tissue wall of a blood vessel or organ, and in particular of the skin, by inhibiting the formation of AGEs; or to prevent and / or fight against the protein glycatton at the tissue level. 4. The use as claimed in any one of the preceding claims, characterized in that said substance is used at a concentration of preferably between 0.001 and 10%. for plant extracts, and between 1.1 CT ⁷ and 1% for the characterized molecules, by weight of the total composition. 5

5. Cosmetic composition, applicable topically or as a dietary supplement, for preventing and / or combating the reduction of the elastic and plastic properties of the skin, by inhibiting the

SUBSTITUTE SHEET (RULE 26) formation of AGEs on the skin, characterized in qu'eue comprises as active substance promoting the inhibition of formation of AGEs fa one substance selected from an extract of a plant _"whole or part thereof, selected from : an extract of a plant, whole or part of it, selected from Epimmede, Qseilfe kinky, Sarsaparilla, liana natives, seeds of Guarana, Cachou, Syfmarin, Pine, Rhubarb of China, Epirnede, Hawthorn , Leucocyanidines, Arquier, Blueberry, Elder, Walnut, Willow, Kinky Leprechaun, Salad, Sarsaparilla, and Lespedeza; and / or a compound chosen from anthraquinone and its derivatives, preferentially 1,4-anthraquinone and 1-arnoin-2-hydroxyrnethylanthraquinone, 4-hydroxychafcone, an OPC (procyansdol oligomers), and OPC strawberry, fes ieucocya nidifies or proanthocyanines of structure (I)

(I), prunin, catechol, 4-aminophenoi _? sodium erythrosine, or a mixture of these substances, optionally in combination with a substance to promote the protection of skin proteins, or a mineralization-inhibiting agent, or an enzyme inhibitor, or a substance capable of

SUBSTITUTE SHEET (RULE 26) to decrease the available sugar level to participate in the gfycation reaction.

6. Composition according to claim 5, in which the active substance is chosen from the group consisting of an epimedal extract (EpiMdium brevicornum), preferably the leaves, a friable osseous extract (Rumex crispus), preferably the root. _The leaves and / or the bark, an extract of sarsaparilla (Smifax orπata), preferably the roots, an extract of vine of the natives (Daviila rugosa), preferentially leaves, the 1,4-anthraquinine, the 4-hydroxycha Icon , OPC strawberry, prunine and a combination of them.

7. Composition according to one of claims 5 to 6, wherein said substance is used at a concentration preferably between 0.001 and 10% for plant extracts, and between 10O ⁷ and 1% for the characterized molecules, by weight of the total composition.

8. Pharmaceutical composition, in particular intended to counteract and / or prevent the reduction of the elastic and plastic properties of a tissue, and in particular of the cutaneous tissue, or of the tissue wall of a blood vessel or of an organ, by inhibition of tissue-forming AGEs, characterized in that the active ingredient promoting the formation of AGEs is a substance selected from an extract of a whole plant or a portion thereof; ci, selected from Epimede, Sorreled Pochard, Sarsaparilla, Liana of Natives, Guarana Seeds, Cachou, Syfmarin, Pine, Rhubarb of China, Epimbus, Hawthorn, Leucocyanidines, Arquier, Blueberry, Elder, Walnut, Willow, Kernel Sorrel, Salad , Sarsaparilla, and Lespedeza, and / or a compound

SUBSTITUTE SHEET (RULE 26) selected from anthraquiπone and its derivatives, preferably 1,4-anthraquinone and 1-amino-2-hydroxymethylanthraquinone, 4-hydroxychalcone, an OPC (procyanidolic oligomers), such as strawberry OPC, leucocyanidines or proanthocyanines structural (I)

(I), prunin, catechol, 4-aminophenol, sodium erythrosine, or a mixture of these substances, for the preparation of a cosmetic, nutraceutical or pharmaceutical composition, or a mixture thereof substances, optionally in combination with a substance to promote protection of tissue proteins, or a glycation-inhibiting agent by mineralization, or an enzyme inhibitor, or a substance capable of decreasing the available sugar level to participate in the reaction of giycatïon.

9. Composition according to claim 7, in which the active substance is chosen from the group consisting of an epimedium extract (Epimedium brevicornum), preferably the leaves, a friable oseiSe extract (Rumex cπspus), preferentially the root, the leaves and / or the bark, an extract of sarsaparilla (Smilax ornata), preferably the roots, a vine extract of the natives (Davilfa rugosa), preferentially

SUBSTITUTE SHEET (RULE 26) leaves, 1,4-anthraquinone, 4-hydroxychalcone, strawberry OPC, prune and a combination of them.

10. Composition according to one of claims 8 to 9, in which said substance is used at a concentration preferably between

0.001 and 10% plant extracts and between 1.10 ^/ 1 and% for the characterized molecules, by weight of total composition fa.

11. A method of screening for active ingredients inhibiting the formation of K) AGEs, comprising; a) contacting at least one type of glycated protein with a substance to be screened for its activity with respect to inhibition of AGE formation; and b) separating at least one active ingredient promoting inhibition of AGE formation.

12. Screening method according to claim 11, characterized in that step a) comprises an incubation of at least one type of protein of the skin or of walls of blood vessels with a reducing sugar or BSA ( bovine serum albumin) with a reducing sugar under conditions allowing the formation of AGEs.

13. The screening method according to claim 11 or 12, characterized in that step b) comprises incubating at least one type of skin protein or blood vessel walls, for example human collagen. , with a reducing sugar or BSA (bovine serum ai) with a reducing sugar in the presence of at least one substance to be screened at

SUBSTITUTE SHEET (RULE 26) a temperature of between 40 and 60 ^° C., preferably about ^{0 °} C.

14. Cosmetic treatment method comprising topical application, or! Ingestion as a dietary supplement, of a composition comprising as cosmetic active ingredient at least one of the ingredients of any one of Claims 1 to 5, or of a cosmetic composition according to one of Claims 7 to 9.

SUBSTITUTE SHEET (RULE 26)