COMPOSITION WITH FLAVANOL AND E-VINIFERIN MONOMERS

Description

Technical field

[0001] The invention relates to a composition comprising the combination of flavanol monomers and ε-viniferine as a nutritional product or medicament intended for humans or animals. State of the art

[0002] The endothelium is a monocellular layer lining the inside of blood vessels, acting as a physical barrier between the tissue layer and the circulating phase. It regulates vascular tone. Dysfunction of endothelial function, or endothelial dysfunction, is characterized by a decrease in the ability of vessels to dilate in response to increased blood flow. It results primarily from a decrease in the availability of nitric oxide (NO), which may be due either to a reduction in NO production or to its increased uptake, particularly due to the phenomenon of oxidative stress.

[0003] Endothelial function is assessed by measuring flow-mediated vasodilation (often referred to as “FMD”).

[0004] The endothelium therefore plays a central role in maintaining blood flow (both at the macro- and microvascular level). Consequently, impaired endothelial function mainly leads to cardiovascular diseases or disorders, particularly atherosclerosis, but also vascular and ocular diseases, diseases associated with cognitive decline, functional intestinal disorders, erectile dysfunction, or disorders associated with menopause.

[0005] Endothelial dysfunction is known to be an early predictive marker of cardiovascular disease, preceding its clinical manifestations. However, cardiovascular disease is the leading cause of death worldwide (30% of deaths worldwide) and represents a major public health problem.

[0006] There is therefore a need to identify a product capable of improving endothelial function and contributing in particular to the prevention of cardiovascular diseases.

[0007] Furthermore, endothelial dysfunction has also been described as a predictive marker of cognitive decline, even in the absence of cerebrovascular disease (Naiberg et al. Psychosom Med. 2016;78(2):192-207). Indeed, cerebral blood flow, which plays a key role in cognitive functions, is closely linked to endothelial function (Fouda et al. Arterioscler Thromb Vasc Biol. 2019 Apr;39(4):593-602).

[0008] Maintaining good vascular health (and therefore optimal endothelial function) is also important for vision. Indeed, cardiovascular diseases (diabetes, hypertension but also hypercholesterolemia) can cause various eye diseases such as hypertensive retinopathy, diabetic retinopathy, glaucoma, etc. (Singh et al. Eur J Ophthalmol. 2020 Apr 27:1120672120914232).

[0009] Endothelial dysfunction is also considered one of the etiological factors of inflammatory bowel diseases (Cibor et al. World J Gastroenterol. 2016 Jan 21;22(3):1067-1077).

[0010] On the other hand, NO, a vasodilator, also plays an essential role in triggering erection. Thus, in men, endothelial dysfunction is often associated with erectile dysfunction (Kovâcs I et al. J.Cardiovasc Pharmacol, 2008 Feb;51(2):148-53).

[0011] In women, estrogens stimulate NO production by endothelial cells. However, during menopause, the drop in estrogen leads to a sharp decrease in FMD, reflecting endothelial dysfunction (Somani YB et al. Am J Physiol Heart Circ Physiol, 2019 Aug 1;317(2):H395-H404). In addition, mental stress has been shown to induce an alteration of endothelial function (Takase B et al. Clin Cardiol. 2004 Apr;27(4):223-7). Similarly, sleep deficit and certain sleep disorders can alter endothelial function (Budhiraja et al. J Clin Sleep Med. 2007 Jun 15;3(4):409-15). Endothelial function, like microcirculation, is also affected by UV exposure (Wolf et al. Exp Physiol. 2019;104(7):1136-1146).

[0012] Thus, endothelial dysfunction is the cause or is involved in many pathologies.

[0013] There is therefore a need to maintain or improve endothelial function.

[0014] Several clinical studies have shown significant improvement in FMD after consuming foods or beverages rich in flavonoids. Among flavonoids, consumption of a single dose of pure epicatechin has been shown to induce a significant increase in FMD within 2 hours (Shafabakhsh et al., Crit Rev Food Sci Nutr. 2020;60(14):2369-2378).

[0015] On the other hand, epidemiological studies have revealed an inverse correlation between dietary flavonoid intake and age-related cognitive decline. In particular, flavanol monomers (catechins and epicatechins) have been shown to exert favorable effects on cognitive functions, and these effects are mainly due to the ability of flavanol monomers to increase cerebral blood flow (Haskell-Ramsay et al., Nutrients. 2018 Jul 27;10(8):986). Studies that evaluated the effects of epicatechin on cognitive functions have shown favorable effects when the dose of epicatechin was more than 50 mg / day, for at least 28 days, in subjects older than 50 years (Haskell-Ramsay et al., Nutrients. 2018 Jul 27;10(8):986).

[0016] Significant increases in FMD (and thus improved endothelial function), improved cerebral blood flow, and some cognitive performance have also been demonstrated after resveratrol supplementation (Cicero et al. Arch Med Sci. 2019;15(4):936-943). Preclinical experiments suggest that resveratrol dimers, particularly ε-viniferine and δ-viniferine, may be able to induce the same effects (Wu et al. The Kaohsiung Journal of Medical Sciences 36 (2020): 535-542).

[0017] Thus, beneficial effects of flavonoids, resveratrol or its dimers individually on endothelial function have already been reported.

[0018] Finally, the prior art also teaches us that Vitis vinifera is a source of flavanol monomers and ε-viniferin (Jayaprakasha et al: "Antioxidant activity of grape seed (Vitis vinifera) extracts on peroxidation models in vitro", May 1, 2001; Gyongyi Nemeth et al: "Stilbenes in the different organs of Vitis vinifera CV. Merlot grafted on TKSBB rootstock", September 26, 2017; US 2018 / 271928; Al-Awwadi Najim a et al: "Extracts enriched in different polyphenolic families normalize increased cardiac NADPH oxidase expression while having differential effects on insulin resistance, hypertension, and cardiac hypertrophy in high-fructose-fed rats" January 12, 2005; FR 3 042 712).

[0019] Uses associated with these molecules alone or in combination are also described, such as in memory (Bensalem Julien et al: "Polyphenols From Grape and Blueberry Improve Episodic Memory in Healthy Elderly with Lower Level of Memory Performance: A Bicentric Double-Blind, Randomized, Placebo-Controlled Clinical Study" June 18, 2019), cognitive disorders (Pierre Philip et al: "Acute Intake of a Grape and Blueberry Polyphenol-Rich Extract Ameliorates Cognitive Performance in Healthy Young Adults During a Sustained Cognitive Effort, December 17, 2019), hyperpigmentation (Yun Cheong-Yong et al: "[alpha]-Viniferin Improves Facial Hyperpigmentation via Accelerating Feedback Termination of cAMP / PKA-Signaled Phosphorylation Circuit in Facultative Melanogenesis" January 1, 2018, Kazuomi Sato et al: of Catechins" November 4, 2009), and as a tyrosinase inhibitor (Zolghadri samaneh et al: "A comprehensive review on tyrosinase inhibitors", January 1, 2019,Likhitwitayawuid et al: “Stilbenes with tyrosinase inhibitory activity”, January 10, 2008).

[0020] However, there is still a significant need for a product capable of preventing or treating endothelial dysfunction and associated diseases, sleep disorders, stress, cognition or even sun protection with improved efficacy compared to the prior art. Summary of the invention

[0021] Also, the objective of the invention is to provide a new composition having greater efficacy than existing products in the fight against diseases associated with endothelial dysfunction compared to said specific molecules consumed individually.

[0022] The inventors have surprisingly identified a composition comprising a specific combination of molecules of natural origin as well as a specific quantity of said molecules present in said composition, exhibiting a synergistic effect and therefore an improved efficiency of endothelial function. They have also demonstrated the interest of said composition for its antioxidant effects, as well as for protecting the skin from UVB (ultraviolet B), preventing sunburn, preventing hyperpigmentation, preventing and / or reducing age-related brown spots. Finally, the composition is also of interest for improving memory and / or attention and / or concentration and / or alertness and / or vigilance and / or learning and / or language and / or mood and / or stress and / or anxiety and / or sleep.

[0023] In particular, the subject of the invention is a composition specifically comprising at least one molecule belonging to the flavonoid family and at least one molecule belonging to the stilbene family. Such a composition is intended to be used as a nutritional product, food supplement or medicine in humans or animals.

[0024] The invention specifically relates to a composition comprising a mixture of molecules comprising: at least 15% of flavanol monomers, the percentage being given by dry weight relative to the total dry weight of the composition, and at least 15 ppm of epsilon-viniferine (ε-viniferine), by dry weight relative to the total dry weight of the composition, said mixture of molecules is obtained from at least one extract or a mixture of extracts of Vitis vinifera, the ratio of flavanol monomers to ε-viniferin is between 20,000 and 4.

[0025] Such a composition thus presents a synergistic effect in comparison with said molecules administered in isolation.

[0026] Thus, the amount of flavanol monomers is greater than the amount of ε-viniferin, namely, the ratio of flavanol monomers to ε-viniferin is between 20000 and 4.

[0027] The flavanol monomers and ε-viniferin are derived from a plant extract, in particular from at least one extract of Vitis vinifera (grape) and / or a mixture of at least two extracts of Vitis vinifera.

[0028] The composition may be administered orally as a nutritional product, dietary supplement, or medicament. The composition and / or one or more molecules of the composition may be encapsulated or microencapsulated in a food carrier. The composition may be in the form of a powder, capsule, tablet, capsule, solution, suspension, emulsion, or chewing gum.

[0029] When the composition is intended to be used as a medicament or dietary supplement, the composition is preferably used to prevent and / or treat diseases chosen from cardiovascular diseases, vascular diseases, diseases associated with cognitive decline, diseases associated with memory loss, neurodegenerative diseases, digestive diseases, joint diseases, erectile diseases and disorders associated with menopause.

[0030] Another aspect of the invention relates to a non-therapeutic use of the composition according to the invention in healthy humans or animals to improve cognitive functions and / or executive functions, and / or to limit normal age-related cognitive decline and / or to improve memory and / or attention and / or concentration and / or alertness and / or vigilance and / or learning and / or language and / or mood and / or stress and / or anxiety and / or sleep and / or to homogenize skin pigmentation, and / or to reduce the appearance of age-related skin spots and / or to regulate melanogenesis and / or to prevent skin hyperpigmentation, and / or to prevent sunburn, and / or to improve the radiance of the complexion.

[0031] Other characteristics and advantages will emerge from the detailed description of the invention, the examples and the figures which follow. Brief description of the Figures

[0032] [ Fig. 1 ] There Figure 1represents, when evaluating tyrosinase activity, the absorbance measurement (normalized and corrected) (mean ±SEM) at 7 min as a function of the different compositions tested as well as their respective inhibition percentages compared to the control. n: number of replicates. Concentrations tested: red grape marc extract: 0.025 mg / ml, licorice extract: 0.0375 mg / ml, grape seed extract: 0.0286 mg / ml, a mixture comprising a composition according to the invention: 0.1 mg / ml. The mixture was composed of 58.8% of a composition according to the invention itself composed of 53.4% ​​of E1 and 46.6% of E3 (the composition according to the invention providing 19.4% of flavanol monomers and 28 ppm of ε-viniferin) and 41.2% of a liquorice extract providing 8.1% of glycyrrhizic acid. p-value for red grape marc extract: <0.724, p-value for liquorice extract: <0.0001, p-value for grape seed extract: <0.0001, p-value for the interaction of the 3 extracts: <0.0001. [. Fig. 2 ] There Figure 2is A: a representation of the % of vasorelaxation observed (mean ± SEM) or estimated as a function of the cumulative volumes of sample added to the organ tank. The curves shown show the effect of a mixture tested at 2 mg / ml, the estimated effect of a seed extract (E1) at 1.1 mg / ml, and the estimated effect of a white grape marc extract (E5) at 0.1 mg / ml. The mixture tested comprises, for its part, 65% of a composition according to the invention, itself composed of 92.3% of E1 and 7.7% of E5 (the composition according to the invention providing 33% of flavanol monomers and 459 ppm of ε-viniferine) and 35% of maltodextrin. The estimated effect of a 1.1 mg / ml seed extract was previously compared with observed data. Statistical analysis by a t-test did not show any significant difference between the estimated values ​​and the observed values ​​for cumulative volumes between 100 µl and 2000 µl.B: A histogram representing the % vasorelaxation (mean ± SEM) for 300µl of samples added to the organ tank. [. Fig. 3 ] There Figure 3 represents the mean fluorescence intensity (MFI) (mean ± SEM) emitted by cells labeled with CM-H2DCFDA. Extract mixture: mixture tested at 270.9 mg / ml and comprising: 53.7% of a composition according to the invention itself composed of 53.4% ​​of E1, and 46.6% of E3 (the composition according to the invention providing 19.4% of flavanol monomer and 28 ppm of ε-viniferine) and 37.5% of a liquorice extract. ***p<0.001. [ Fig. 4 ] There Figure 4represents the keratinocyte proliferation rate expressed in % (reduced to the non-irradiated control for each condition, mean ± SEM). Extract mixture: mixture tested at 270.9 mg / ml and comprising: 53.7% of a composition according to the invention itself composed of 53.4% ​​of E1, and 46.6% of E3 (the composition according to the invention providing 19.4% of flavanol monomers and 28 ppm of ε-viniferine) and 37.5% of a liquorice extract. ***p<0.001. Detailed description of the invention Definition

[0033] For the purposes of the invention, “animal” means any animal excluding humans.

[0034] By "extract from Vitis vinifera » or « grape extract » within the meaning of the invention means an extract comprising at least one molecule, in particular at least flavanol monomers and / or ε-viniferin, preferably a set of molecules, obtained from Vitis vinifera and / or a by-product of Vitis vinifera.The raw material may be the leaves and / or the fruits and / or the seeds and / or the skins and / or the stalks and / or the wood parts (shoots, vines, roots), and / or the marc and / or the wine, preferably the raw material is the grape marc (including the seeds, the skins and possibly the stalks) and / or the grape seed.

[0035] By "ppm" we mean parts per million in the mixture.

[0036] For the purposes of the invention, the term "nutritional product" means a food ingredient for nutritional purposes used alone or in combination with other ingredients or food additives in food formulas, including food supplements and foods intended for humans or animals.

[0037] For the purposes of the invention, the term "medicine" means an active ingredient used for therapeutic purposes, used alone or in combination with other active or non-active substances in medicinal formulas, including herbal medicine or food supplements with a therapeutic effect, intended for humans or animals.

[0038] By "prevent" or "prevention" within the meaning of the invention, we mean the reduction to a lesser degree of the risk or probability of occurrence of a given phenomenon, that is to say, in the context of the present invention of a disease or a disorder.

[0039] For the purposes of the invention, the term "treat" or "treatment" means a reduction in the progression of the disease or disorder, a stabilization, a reversal or regression, or even an interruption or inhibition of the progression of a disease or disorder. In the context of the invention, these terms also apply to one or more symptoms of said diseases or disorders of the present invention. Composition

[0040] The present invention therefore relates to a new composition exhibiting in particular increased efficacy in improving or maintaining endothelial dysfunction and in sun protection.

[0041] To this end, the invention relates to a composition comprising a mixture of molecules comprising: at least 15% of flavanol monomers, the percentage being given by dry weight relative to the total dry weight of the composition, and at least 15 ppm of ε-viniferine, by dry weight relative to the total dry weight of the composition, said mixture of molecules is obtained from at least one extract or a mixture of extracts of Vitis vinifera, the ratio of flavanol monomers to ε-viniferin is between 20,000 and 4.

[0042] Flavanol monomers are compounds of the flavonoid family, in particular of the flavanol subclass including in particular flavan-3-ols or flavanols or catechins. The most abundant class is made up of catechins and / or epicatechins. These are powerful antioxidants which help in particular to prevent inflammatory and coronary diseases but also to maintain or increase cognitive performance. In the context of the invention, the flavanol monomers are chosen from catechins, epicatechins, as well as their galloylated forms, such as epicatechin-3-O-gallate, epigallocatechin, epicatechin gallate, epigallocatechin gallate. These monomers can be extracted from one or more plants (in whole or in part) such as tea (Camelia sinensis), apples (Malus domestica), cocoa (Theobroma cacao) or from plants (in whole or in part) belonging to the genus Vitis and preferentially Vitis vinifera.They can also be derived from microalgae. ε-viniferin is a natural phenol from the stilbene family, particularly stilbenoids. ε-viniferin is also a dimer of resveratrol. It can be extracted from one or more plants such as plants (whole or part) belonging to the genus Iris - Iridaceae; Sophora - Fabaceae; Gnetum - Gnetaceae; Carex - Cyperaceae; Peony (Paeonia) - Paeoniaceae; Dipterocarpus - Dipterocarpaceae and more particularly belonging to the genus Vitis, preferentially Vitis vinifera. ε-viniferine can also be derived from microalgae.

[0043] According to an object of the invention, the composition preferably comprises at least 100 ppm of stilbenes comprising said at least 15 ppm of ε-viniferine. Thus, the composition according to the invention comprises at least 100 ppm of stilbenes including at least 15 ppm of ε-viniferine. In other words, the composition may comprise for example 85 ppm of stilbenes other than ε-viniferine and 15 ppm of ε-viniferine. Said other stilbenes may be pinosylvin, piceatannol, trans-resveratrol, trans-pterostilbene, rhapontigenin, isorhapontigenin, rhapontine, ponticine, trans-piceide, or astringine. (delta and / or omega and / or alpha and / or R and / or R2) viniferine, ampelosin A and / orE and / or F and / or H, miyabenol C, parthenocissine A, pallidol, vitisine C, hopeaphenol, isohopeaphenol, viniferol E.

[0044] According to a preferred embodiment, the composition according to the invention may comprise at least 20% of flavanol monomers, the percentage being given in dry weight relative to the total dry weight of the composition, more preferably at least 25% of flavanol monomers.

[0045] According to another embodiment, the composition according to any of the preceding embodiments may comprise at least 50 ppm of ε-viniferine, by dry weight relative to the total dry weight of the composition, more preferably at least 100 ppm of ε-viniferine.

[0046] The inventors have identified that the composition according to the invention exhibits improved activity when the amount by weight of flavanol monomers is greater than the amount by weight of stilbene, preferably ε-viniferine.

[0047] The ratio of flavanol monomers to ε-viniferin is thus between 20,000 and 4. By ratio is meant the ratio of magnitude between the quantity of flavanols and the quantity of ε-viniferin (flavanol monomers / ε-viniferin). In particular, the ratio is between 15,000 and 30, even more preferably between 10,000 and 100.

[0048] Flavanol monomers are of natural origin. These molecules can thus be obtained from a natural product, namely a plant extract chosen from an extract of Vitis, preferentially Vitis vinifera, a tea extract (Camelia sinensis), an apple extract ( Malus domestica ), a cocoa extract (Theobroma cacao). They can also be derived from microalgae.

[0049] ε-viniferine is of natural origin and can thus be obtained from a natural product, namely a plant extract chosen from a Vitis extract, preferably Vitis vinifera.

[0050] When the flavanol monomers and ε-viniferin are obtained from the same plant, the plant is a Vitis extract, in particular an extract of Vitis vinifera.

[0051] It is already known that grapes and wine contain flavanol monomers. However, the presence of stilbenes and in particular ε-viniferin is not systematic, as shown in Table 1 below. In addition, the monomer concentration of Vitis vinifera never naturally exceeds 10% of total polyphenols.

[0052] However, the composition according to the invention aims at a mixture of molecules comprising at least 15% of flavanol monomers. Thus, the origin and type of the raw material allowing the extraction of the extract(s) as well as the process applied allowing the extraction thereof are combined in order to obtain a mixture of molecules comprising at least 15% of flavanol monomers and at least 15 ppm of ε-viniferine. The concentrations of molecules according to different grape raw materials are given in Table 1 below. [Table 1] Flavanol monomers stilbenes Vitis vinifera Total Polyphenols (TP) (Folin) Catechin (C) Epicatechi ne (EC) Total C+EC [C+EC] / PT Including Trans-resveratrol Including ε-viniferine Grape Juice (mg / 100ml) / (%) or ppm 1.21 1.26 2.47 (0.002%) 0.0508ppm - Black grape (mg / 100g) / (%) or ppm 184.97 (0.18%) 5.46 5.24 10.7 (0.01%) 5.8% 1.5 ppm - White grape (mg / 100g) / (%) or ppm 121.80 (0.12%) 1.41 0.49 1.90 (0.002%) 1.6% 0.3 ppm - Red wine (mg / 100m) (%) or ppm 215.48 (0.22%) 6.81 3.78 10.59 (0.01%) 4.9% 1.8 ppm 1.5 ppm Rosé wine (mg / 100m) / (%) or ppm 82.21 (0.08%) 0.91 0.55 1.46 (0.001%) 1.8% 0.6 ppm - White wine (mg / 100ml) / (%) or ppm 32.10 (0.03%) 1.08 0.95 2.03 (0.002%) 6.3% 0.3 ppm 0.0557pp m Invention >50 % >15% >30% 15 ppm

[0053] According to a preferred embodiment, the extract(s) of Vitis vinifera in the composition according to the invention when it contains, comprise(s) at least 50% of total polyphenols, the percentage being given in dry weight relative to the total weight of the extract of Vitis vinifera.

[0054] Preferably, the extract of Vitis vinifera is an extract of grape marc and / or grape seed, more preferably the composition comprises at least one grape seed extract and / or one grape marc extract. According to one embodiment, the composition according to the invention comprises at least one grape seed extract and one grape marc extract.

[0055] In addition to flavanol monomers and ε-viniferine, the composition according to the invention may comprise dimers (PAC B1 and B2). Preferably, the concentration of flavanol dimers (PAC B1 and B2) is greater than 5%, the percentage being given in dry weight relative to the total dry weight of the composition.

[0056] When the composition according to the invention comprises at least one grape marc extract, preferably the grape marc extract(s) preferably comprise flavonols. Preferably, the concentration of flavonols in the composition is greater than or equal to 0.05% by dry weight relative to the total dry weight of the composition.

[0057] The composition according to the invention may comprise, in addition to the mixture of molecules described above, at least one other constituent chosen from antioxidants, natural extracts, vitamins, trace elements, carotenoids, omega 3 fatty acids, natural oils, and mixtures thereof.

[0058] When the composition according to the invention further comprises a natural extract other than the extract of Vitis vinifera, the natural extract is preferably a licorice extract.

[0059] Licorice is used in traditional medicines, with the earliest recorded medicinal uses dating back to ancient Assyrian, Egyptian, Chinese, and Indian cultures.

[0060] The benefits attributed to it are numerous. When taken orally, licorice was used in remedies for disorders of the respiratory, gastrointestinal, cardiovascular and genitourinary systems. Topically, it was used for the treatment of certain eye or skin diseases (Fiore et al. J Ethnopharmacol 2005 Jul 14;99(3):317-24). External use, particularly topical, is also documented. Thus, licorice extract is often included in cosmetic formulations aimed at improving the appearance of the skin.

[0061] Preferably, the licorice extract comprises glycyrrhizic acid and / or glabridin.

[0062] As an example of additional plant extracts, the composition preferably also comprises at least one extract chosen from extracts of coffee, olive leaves, blackcurrant, polygonum, citrus fruits, marigold and mixtures thereof.

[0063] The polyphenol(s) possibly present in the composition according to the invention may be chosen, for example, from polyphenols, in particular in the form of plant extracts (extracts of olive leaves or olives, blackcurrant, coffee, polygonum, citrus fruits, capers), or vitamin C, in particular in the form of plant extracts (acerola extract, pomegranate extract, citrus fruits), vitamin E, vitamin A, in particular in the form of plant extracts; or their derivatives, zinc, selenium and their mixtures.

[0064] The vitamin(s) optionally present in the composition according to the invention may be chosen, for example, from vitamin B9, B12, C, D, E, A including pro-vitamins A, their derivatives and their mixtures.

[0065] The trace element(s) possibly present in the composition according to the invention may be chosen, for example, from zinc, selenium or chromium and their mixtures.

[0066] Zinc is an essential mineral for proper brain function by improving insulin sensitivity, reducing oxidative stress and inflammation. Moreover, the European Food Safety Authority, under Regulation 1924 / 2006, recognizes the role of zinc in normal cognitive function.

[0067] The carotenoid(s) optionally present in the composition according to the invention may be chosen, for example, from lutein, zeaxanthin, crocin, picrocrocin and mixtures thereof.

[0068] The moga-3 fatty acid(s) optionally present in the composition according to the invention may be chosen, for example, from docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) and mixtures thereof.

[0069] Long-chain omega-3 fatty acids such as docosahexaenoic acid (DHA) have been shown to have a beneficial effect on cognitive function at the preclinical level in aged mice. Its beneficial effects on neuroinflammation are now well accepted and widely described in the literature.

[0070] The natural oil(s) possibly present in the composition according to the invention may be chosen, for example, from fish oils, vegetable oils, oils from microalgae and their mixtures.

[0071] The composition may also include other constituents, such as excipients, coating agents such as maltodextrin, microcrystalline cellulose, cyclodextrins, starch, soluble or insoluble fibers, water or alcohol.

[0072] According to another subject of the invention, the composition and / or one or more molecules of the composition may be encapsulated or microencapsulated in a food carrier chosen from maltodextrin, gum arabic, hydrogenated oil, non-hydrogenated oil, wax, alginate, starch, protein and mixtures thereof.

[0073] Encapsulation is particularly advantageous and makes it possible to stabilize and protect the mixture of molecules according to the invention from its environment, in particular from the human body or the animal that ingests the composition. Thus, the flavanol monomers and ε-viniferine are protected and their bioavailability is improved.

[0074] In addition, encapsulation allows the trapping of volatile and thermosensitive active metabolites in the matrix, improving the stability, bioavailability and use of said mixture of molecules in food matrices, including masking of taste and resistance to possible alterations of the compounds during the production stages of a food product.

[0075] According to one variant, the composition and / or one or more molecules may be microencapsulated. Microencapsulation makes it possible to protect the active substances within particles of sizes between 1 µm and 1000 µm, more preferably between 30 µm and 500 µm and even more preferably between 50 µm and 300 µm.

[0076] According to another object, the composition according to the invention is in the form of a powder, capsule, tablet, capsule, solution, suspension, emulsion or chewing gum.

[0077] Where the composition is intended for use as a nutritional product, food product or food supplement, the composition may be in the form of a dairy product, cereal, grain product or beverage.

[0078] Preferably, the composition according to the invention is delivered at a dose making it possible to provide humans or animals daily with at least 0.25 mg per kg of body weight of flavanol monomers and 0.025 µg per kg of body weight of ε-viniferine. Process

[0079] The composition according to the invention can be obtained by any process making it possible to obtain a mixture of molecules comprising at least 15% of flavanol monomers by weight relative to the total weight of the composition and at least 15 ppm of ε-viniferine by weight relative to the total weight of the composition. This may be the mixture of the different molecules in the desired proportions or the mixture of plant extracts comprising the different molecules in the desired proportions.

[0080] According to one embodiment, the composition according to the invention is obtained from an extract of Vitis vinifera or a mixture of extracts of Vitis vinifera. The extracts may be obtained by any process which makes it possible to obtain a mixture comprising at least 15% of flavanol monomers by weight and at least 15 ppm of ε-viniferin by weight.

[0081] A process particularly suitable for obtaining an extract of Vitis viniferaincludes the following steps: extraction with water and / or alcohol (especially ethanol) of Vitis vinifera (e.g. fruit, skin, seed, leaf, stalk, vine shoot, vine, wood, root), preferably of seeds of Vitis vinifera and / or marc de Vitis vinifera (the marc containing grape skins, i.e. grape skin, grape seeds and possibly stalks), the seeds and / or marc may come in particular from pressing residues from wine production, and / or from grape leaves Vitis vinifera and / or vine shoots, vines, wood, roots of Vitis vinifera.The quantity of water or hydroalcoholic solution (30% v / v to 96% V / V) used is preferably between 2 and 10 times the mass of material used. The extraction time can be between 30 minutes and 24 hours and the extraction temperature between 20°C and 80°C. The raw materials used can be in dry, fresh, or frozen whole or ground forms; separation of the water and / or alcohol solution from the solid matter, for example by centrifugal decantation or by pressing and filtration; the extract thus obtained is called crude extract A, possibly an elimination of the alcohol by vacuum evaporation at a temperature, preferably lower than 60°C and at a pressure lower than 100mbars.

[0082] If more than one raw material is used, for example grape seeds and grape marc, the process may involve a single extraction from the two raw materials mixed together or the process may involve obtaining two separate extracts (implementing all or part of the process separately for each extract, then mixing the two extracts obtained at the end).

[0083] Based on the crude extract(s) A previously obtained, the process may comprise a specific step to obtain one or more purified extract(s) rich in flavanol monomers and / or in viniferine.

[0084] For flavanol monomers, purification can be carried out either: by purification by ultrafiltration and / or nanofiltration membrane separation known to those skilled in the art of the extract previously desolventized so as to preferentially select the monomers and eliminate the flavanol polymers, to obtain an extract comprising a fraction enriched in flavanol monomers, by purification using a liquid / liquid separation by organic solvents known to those skilled in the art and in particular by ethyl acetate, by purification via precipitation with salts such as NaCl of the flavanol polymers followed by a solid / liquid separation such as filtration for example.

[0085] The extract thus obtained is a purified extract rich in flavanol monomers called extract B.

[0086] The process preferably comprises an additional step of chromatographic purification of extract A and / or extract B. This step consists of passing extracts A and / or B through a chromatographic column containing adsorption resins particularly suited to polyphenols and known to those skilled in the art, then rinsing the resins using an aqueous solution followed by an elution phase using a hydroethanolic solution preferably between 60 and 80% v / v.

[0087] The alcohol of the eluates thus obtained is evaporated and the eluate concentrated via a vacuum concentration step of the eluates rich in polyphenols and more particularly in flavanol monomers and / or in ε-viniferine at a temperature preferably lower than 60°C and a pressure lower than 100 mbar.

[0088] The process may include an additional step which consists of drying the purified extracts by techniques known to those skilled in the art such as atomization, drying in a vacuum oven or drying by lyophilization with or without a support such as maltodextrin, gum arabic for example.

[0089] The method may also comprise a step of encapsulation or microencapsulation of the molecules and / or the composition.

[0090] Once the extract has been obtained, the process for preparing the composition according to the invention may comprise mixing extracts of Vitis vinifera rich on the one hand in flavanol monomers and on the other hand in ε-viniferine. Use

[0091] According to another aspect, the invention also relates to the composition according to any of the preceding embodiments for its use as a medicament for humans or animals.

[0092] Preferably, the composition according to the invention is intended to be used in the prevention and / or treatment of diseases associated with endothelial dysfunction chosen from cardiovascular diseases, vascular diseases, diseases associated with cognitive decline, diseases associated with memory loss, neurodegenerative diseases, digestive diseases, joint diseases, eye diseases, erectile disorders and disorders associated with menopause.

[0093] When the composition is intended to prevent and / or treat diseases associated with cognitive decline including neurodegenerative diseases or diseases associated with memory loss, said disease is chosen from Alzheimer's disease, Parkinson's disease, Huntington's disease, motor neuron diseases, dementia, depression, anxiety, schizophrenia, mental retardation and cognitive dysfunction syndrome (CDS).

[0094] When the composition is intended to prevent and / or treat cardiovascular diseases, said disease is chosen from high blood pressure, atherosclerosis, myocardial infarction, angina pectoris (angina)

[0095] When the composition is intended to prevent and / or treat vascular diseases, said disease is chosen from chronic venous insufficiency, venous thrombosis, varicose veins.

[0096] When the composition is intended to prevent and / or treat digestive diseases, said disease is chosen from irritable bowel syndrome, Crohn's disease, ulcerative colitis and dyspepsia.

[0097] When the composition is intended to prevent and / or treat joint diseases, said disease is chosen from osteoarthritis, rheumatoid arthritis, rheumatoid polyarthritis and ankylosing spondylitis.

[0098] When the composition is intended to prevent and / or treat eye diseases, said disease is chosen from age-related macular degeneration (AMD), hypertensive retinopathy, diabetic retinopathy, glaucoma, cataract or Fuchs' endothelial dystrophy.

[0099] When the composition is intended to prevent and / or treat sleep disorders, said disorder is chosen from insomnia, parasomnia, sleep apnea and / or hypopnea.

[0100] When the composition is intended to prevent and / or treat disorders associated with menopause, said disorder is chosen from hot flashes, night sweats, increased abdominal fat mass, vaginal dryness, decreased libido, mood disorders.

[0101] According to another aspect, the invention also relates to a non-therapeutic use of the composition according to the invention in healthy humans or animals, to improve cognitive functions and / or executive functions, and / or to limit normal age-related cognitive decline and / or to improve memory and / or attention and / or concentration and / or alertness and / or vigilance and / or learning and / or language and / or mood and / or stress and / or anxiety and / or sleep and / or to homogenize skin pigmentation, and / or to improve the beauty of the skin and / or to reduce the appearance of age-related skin spots, and / or to regulate melanogenesis and / or hyperpigmentation due to overproduction of melanin and / or to prevent skin damage and / or disorders linked to sun exposure such as erythema and skin aging.

[0102] When the composition according to the invention is intended to be used to homogenize the pigmentation of the skin, and / or improve the beauty of the skin and / or reduce the appearance of age-related skin spots, and / or regulate melanogenesis, and / or hyperpigmentations due to overproduction of melanin, and / or the prevention of skin damage and / or disorders linked to sun exposure such as erythema and skin aging, the composition preferably comprises at least one liquorice extract.

[0103] By hyperpigmentations due to an overproduction of melanin, we mean, within the meaning of the invention, post-inflammatory hyperpigmentation, or due to phytophotodermatitis, a neoplastic process (lentiligo, melanoma), melasma, freckles, acanthosis nigricans, resulting from taking medication or sun exposure.

[0104] The invention is now illustrated by non-limiting examples of compositions according to the invention, of extract according to the invention, of extract outside the invention and by efficacy results. Examples Example 1 : Grape seed extract rich in flavanol monomers.

[0105] The extract is obtained according to the following process: 40 kg of dried grape seeds aqueous extraction in 400 kg of water heated to 85°C Liquid / solid separation then the aqueous extract is then passed through a membrane < 15kD The permeate is then passed through a C18 adsorption chromatographic column Purification on adsorption resin 10 BV at 2 BV / h, Water washing 4 BV at 2BV / h Hydro-ethanolic elution with 80% V / V solution at 1BV / H with 2BV Concentration under vacuum and dealcoholization under 60 mbar at 40°C Spray drying

[0106] The grape seed extract thus obtained comprises at least 15% flavanol monomers and comprising at least 50% total polyphenols, measured by the Folin method. Such an extract (E1) is characterized in Table 2 below. Example 2: Red grape skin extract rich in anthocyanins but without stilbenes.

[0107] The extract is obtained according to the following process: 40kg of red grape marc Extraction with 200kg Aqueous solution with 0.1% sulfites Concentration under vacuum 100mbar at 50°C partial elimination of sulfites Spray drying.

[0108] The grape extract thus obtained (E2) is characterized in Table 2 below. Example 3: Grape marc extract including stilbenes.

[0109] The extract is obtained according to the following process: Hydroalcoholic extraction Red grape marc extract (E3) 10kg of dry marc hydroalcoholic extraction at 30% V / V solid liquid ratio 1 / 10 stirring at 350 rpm at 80°C for 4 hours Solid liquid separation by filtration at 20µm Vacuum concentration at 40°C Spray drying

[0110] The grape extract thus obtained (E3) is characterized in Table 2 below. Example 4 : Purified red grape marc extract including stilbenes.

[0111] The extract is obtained according to the following process: Extraction 80°C for 4H at 50% v / v EtOH ratio 1 / 10 10kg of dry marc hydroalcoholic extraction at 30% V / V solid liquid ratio 1 / 10 stirring at 350 rpm at 80°C for 4 hours Solid liquid separation by filtration at 20µm Concentration under vacuum at 40°C Purification on adsorption resin 20 BV at 2 BV / h Washing with water 4 BV at 2BV / h Hydroethanolic elution with 80% V / V solution at 1BV / H with 2BV Concentration under vacuum and dealcoholization under 200 mbar at 40°C Drying by lyophilization

[0112] The grape extract thus obtained (E4) is characterized in Table 2 below. Example 5 : White grape marc extract including stilbenes.

[0113] The extract is obtained according to the following process: 125kg of white grape marc hydroalcoholic extraction at 30% V / V solid liquid ratio 1 / 10 stirring at 350 rpm at 80°C for 4 hours Solid liquid separation by filtration at 20µm Concentration / dealcoholization under vacuum under 100 mbar at 40°C Purification on adsorption resin 20 BV at 2 BV / h Washing with water 4 BV at 2BV / h Hydroethanolic elution with 80% V / V solution at 1BV / H with 2BV Concentration under vacuum and dealcoholization under 200 mbar at 40°C Spray drying.

[0114] The grape extract thus obtained (E5) is characterized in Table 2 below. Example 6: Vine shoot extract including stilbenes but without flavanol monomers.

[0115] The extract is obtained according to the following process: 40g of vine shoots hydroalcoholic extraction in 250g of a 90%V / V solution at 80°C for 4 hours Solid liquid separation filtration Dealcoholization and Concentration under vacuum Drying by lyophilization.

[0116] The extract from Vitis viniferaobtained (E6) includes 5.1% resveratrol, 8.5% ε-viniferine, 15.5% stilbenes and 0% catechin and epicatechin Example 7: Examples of compositions according to the invention

[0117] Table 3 below presents several examples of compositions comprising extracts from the processes described previously in Examples 1 to 6. [Table 3] Examples according to the invention Formulation Monomers of flavan ols Dimers (PAC B2 and B1) of flavanols E-viniferi ne Stilbenes Monomers / ε-viniferine ratio Flavonols P Total polyphenols A 75%E1+ 25%E3 26.9% 10,0% 0.0015 % 0.0144% 17639 / 1 0.11% 83% B 75%E1+ 25%E4 29.1% 10.6% 0.0132 % 0.0434% 2200 / 1 0.16% 74% C 99%E1+ 1%E5 35.3% 9.8% 0.0060 % 0.0112% 6272 / 1 0.05% 99% D 40%E1+ 60%E5 15.5% 5.3% 0.3577 % 0.6713% 43 / 1 0.34% 82% E 40%E1+ 35%E2+ 25%E3 15.5% 5.4% 0.0015 % 0.0144% 10160 / 1 0.11% 60% F 50%E1+ 50%E6 17.8% 6.6% 4.2800 % 8.8700% 4.15 / 1 - 76% It should be noted that the concentration of dimers (PAC B1 and B2) of flavanol is greater than 5% in dry weight of the weight of the composition. In addition, for extracts of Vitis vinifera from the parts of the marc (seed, skin, stalk) the flavonol concentration is equal to or greater than 0.05% in dry weight of the weight of the composition. Example 8: Evaluation of the effect of the composition according to the invention on the activity of tyrosinase. Evaluation of the effect of a composition comprising flavanol monomers and ε-viniferin

[0118] The objective of this test is to model the activity of an enzyme, tyrosinase (enzyme responsible for melanin formation) using an experimental design and to determine the influence of flavanol monomers and ε viniferine on the enzymatic activity towards a specific substrate (L-DOPA). Tyrosinase activity was measured in vitro by a colorimetric reaction. Under the action of tyrosinase, L-DOPA is oxidized and converted into dopaquinone. The formation of dopaquinone can then be monitored by spectrometry at 475nm.

[0119] In a 96-well plate, the following quantities were deposited in each well: 80µl phosphate buffer (50mM), 40µl of test sample or buffer, 40µl of tyrosinase at 125U / ml. At the last moment, 40µl of L-DOPA (2.5mM) was added to initiate the reaction and then the absorbance at 475nm was monitored for 25min.

[0120] For each well, the absorbance values ​​over time (A x ) are normalized with respect to the first absorbance measurement (A TO ) according to the following formula: Normalized absorbance (A n ) = A x - A TO . The normalized absorbances are then corrected (for each time) according to the following formula: Corrected normalized absorbance (A nc ) = A n - A CLT where A CLT is the value of the normalized absorbance of the reagents in the presence of the sample (buffer + tyrosinase + sample). The lower the values ​​(A nc ) observed, the more the tyrosinase activity is reduced (with respect to the substrate L-DOPA).

[0121] To carry out this test, expert design software was used, and a centered composite design as the experimental design with for the factors, concentrations between 1 and 50µM for flavanol monomers and between 0.0050 and 0.05µM for ε-viniferine and for the response, the corrected normalized absorbance measured after 7 min. The ratio of flavanol monomers and ε-viniferine is between 10000 and 1000.

[0122] The results obtained showed that the corrected normalized absorbance could be modeled by a quadratic linear model (p<0.0001) (Table 4). [Table 4] Source Sum of squares df Root mean square F-value p-value Model 0.0349 5 0.0070 27.53 < 0.0001 A-Flavanols 0.0275 1 0.0275 108.46 < 0.0001 B-ε-Viniferin 0.0005 1 0.0005 1.88 0.1780 AB 0.0023 1 0.0023 9.00 0.0047 A 2< 0.0030 1 0.0030 11.76 0.0015 B 2< 0.0006 1 0.0006 2.42 0.1285 Residual 0.0096 38 0.0003 Lack of Fit 0.0019 3 0.0006 2.86 0.0509 Value R 2< 0.7837 Adjusted R 2< 0.7552 Predicted R 2< 0.7025

[0123] The results also demonstrate a significant effect of flavanol monomers on tyrosinase activity toward the substrate L-DOPA. Analysis of the coefficients in terms of coded factors (Table 5) shows that flavanol monomers decrease tyrosinase activity toward its substrate, L-DOPA. [Table 5] Postman Estimated Coefficient df Standard error 95% CI (low) 95% CI (high) Intercept 0.1508 1 0.0041 0.1426 0.1591 A-Flavanols -0.0338 1 0.0032 -0.0404 -0.0273 B-ε-Viniferin -0.0045 1 0.0032 -0.0110 -0.0021 AB -< 0.0119 1 0.0040 -0.0200 -0.0039 A 2< 0.0171 1 0.0050 0.0070 0.0273 B 2< 0.0078 1 0.0050 -0.0024 0.0179

[0124] In contrast, no significant effect of ε-viniferin was observed in the concentration range tested in the experimental design. Finally, interestingly, a significant interaction between flavanol monomers and ε-viniferin was observed. Moreover, the coefficient of the interaction of these factors being negative, this demonstrates that the association of flavanol monomers and ε-viniferin also decreases the activity of tyrosinase towards the substrate L-DOPA.

[0125] The effect of the combination of flavanol monomers and ε-viniferine is greater than an additive effect of the molecules. Indeed, for a concentration of flavanol monomers at 35µM and a concentration of ε-viniferine at 0.005µM, an absorbance value of 0.158 is obtained while the absorbance value for an additive effect is 0.332. Therefore, the composition according to the invention does indeed have a synergistic effect. Evaluation of the effect of a mixture of extracts of Vitis vinifera and licorice extract.

[0126] The objective of this test is to evaluate the activity of tyrosinase (with respect to the substrate L-DOPA) in the presence of a mixture comprising: 58.8% of a composition according to the invention itself composed of 53.4% ​​of E1 and 46.6% of E3 (the composition according to the invention providing 19.4% of flavanol monomers and 28 ppm of ε-viniferine) and 41.2% of a liquorice extract providing 8.1% of glycyrrhizic acid. This activity will be compared to the enzymatic activities in the presence of E1 or E3 or licorice extract alone.

[0127] To do this, the activity of tyrosinase was monitored and measured in vitro by the colorimetric reaction described previously. The extracts were tested at the following concentrations: grape seed extract at 0.028 mg / ml, red grape marc extract at 0.025 mg / ml, licorice extract at 0.037 mg / ml.

[0128] The percentage of tyrosinase inhibition was determined on the normalized and corrected absorbances, at T=7min, according to the following equation. % d ′ inhibition = A nc Contrôle − A nc Echantillon A nc Contrôle × 100

[0129] Where A nc Control is the average of the normalized and corrected absorbance measurements of the normal reactions (buffer + tyrosinase + L-DOPA) and A nc Sample is the corrected normalized absorbance of the reaction in the presence of the sample (buffer + tyrosinase + L-DOPA + sample).

[0130] Absorbance values ​​(at T=7min) were statistically compared using a three-way analysis of variance (ANOVA) where factor 1 was treatment with grape seed extract, factor 2 was treatment with red grape pomace extract, factor 3 was treatment with licorice extract. A value of p<0.05 was considered significant.

[0131] In the presence of red grape marc extract, a small and non-significant decrease in tyrosinase activity was observed ( Figure 1). In the presence of a licorice extract or a grape seed extract, significant decreases of 40.2% and 49.8% respectively in tyrosinase activity were observed. Finally, a significant interaction was found between a seed extract, a red grape marc extract and a licorice extract. Thus, such a mixture according to the invention showed a decrease (84.9%) in tyrosinase activity greater than that observed for red grape marc extracts, a grape seed extract or a licorice extract alone. The decrease in tyrosinase activity with respect to the L-DOPA substrate could thus allow the reduction of melanin production and consequently improve skin homogeneity and reduce age-related pigmentation spots or hyperpigmentation processes. Example 9: Evaluation of the effect of a composition according to the invention on endothelial function.

[0132] This test aims to evaluate the vasorelaxation, ex vivo, of rat aortas, in the presence of a mixture comprising: 65% of a composition according to the invention itself composed of 92.3% of E1 and 7.7% of E5 (the composition according to the invention providing 33% of flavanol monomers and 459 ppm of ε-viniferine, and 35% of maltodextrin The vasorelaxation induced by this mixture will be compared to the vasoralaxation induced by extracts E1 and E5 alone.

[0133] To do this, rat thoracic aortas were harvested, cleaned, and sectioned into 2-3mm rings. The rings were then immersed in an isolated organ tank, in 20ml of Krebs solution (118 mM NaCl, 25 mM NaHCO3, 5.5 mM d-glucose, 4.7 mM KCl, 1.18 mM KH2PO4, 2.4 mM MgSO4, and 3.3 mM CaCl2, pH 7.8) at 40°C and continuously bubbled with 95% O2 and 5% CO2. To test endothelial reactivity, contractions were induced with KCl (80mM) until a plateau was reached. The organ tank was then washed 3 times with Krebs solution. Functional endothelium was then tested by first adding 0.5% norepinephrine (at 0.1 µM), to induce contraction, and then acetylcholine (at 10 µM), to induce at least 80% vasorelaxation. Cumulative volumes of sample were then added to the organ vessel to reach a final volume of 2000 µL over a period of 1 h and a concentration-relaxation curve was constructed.Three independent tests were performed for each sample.

[0134] The results show that the mixture comprising a composition according to the invention induces strong vasorelaxation of the aorta sections ( Figure 2A ). Indeed, the maximum vasorelaxation observed is 58% and is reached after adding 300µL of sample. In the presence of the mixture comprising a composition according to the invention, the effect on vasorelaxation is greater than that observed for a grape seed extract alone (and at the same concentration as in the mixture comprising a composition according to the invention) and that estimated for a white grape marc extract alone (and at the same concentration as in the mixture comprising a composition according to the invention) ( Figure 2B). The mixture comprising a composition according to the invention also induces an effect greater than the addition of the effects of the extracts (outside the invention) taken separately. These results demonstrate that there are synergistic interactions between the flavanol monomers and ε-viniferine in the mixture comprising a composition according to the invention. Such a synergistic effect of the composition of the invention makes it possible to propose formulations improving endothelial function and therefore vascular function. Example 10: Evaluation of the antioxidant capacity of a mixture of extracts of Vitis vinifera

[0135] The objective of this test is to determine the antioxidant activity of extracts of Vitis vinifera such as grape seed extract or a mixture of extracts from Vitis viniferacomprising 50% E1 and 50% E3 by comparing it to a vitamin E analogue: trolox. The antioxidant activity was measured, spectrophotometrically, by the oxygen radical absorbing capacity (ORAC) test. This test is based on the fact that the fluorescence of the fluorescein molecule decreases under the action of an oxidizing agent: AAPH (“2,2'-Azobis(2-amidinopropane) dihydrochloride”), which generates peroxyl radicals. The presence of antioxidants prevents or slows down the loss of fluorescence, measured, which is calculated by the area under the curve as a function of time. The antioxidant capacity of the tested sample is then expressed in Trolox equivalent, i.e. as a function of the concentration of Trolox having the same activity as the tested sample, at a given concentration. The ORAC value obtained is independent of the concentration tested for the sample.

[0136] In a 96-well plate, the following quantities were placed in each well: 25µl of phosphate buffer (50mM, pH 7.5) for blanks or 25µL of Trolox (25µM, 20µM, 15µM, 10µM, 5µM, 2µM), 25µL of diluted sample, 150µL of Fluorescein (8nM). The plate was then incubated for 30 min at 37°C. Just before reading, 25µl of AAPH (153mM) was added.

[0137] The results show that grape seed extracts as well as the mixture of grape seed extracts Vitis vinifera have strong antioxidant capacity with ORAC values ​​between 10524 and 24142. Thus a mixture of extracts of Vitis vinifera according to the invention makes it possible to offer formulations with a high antioxidant power. Given that it is recommended to consume 3000 to 5000 ORAC units each day in order to reduce oxidative stress, such a composition according to the invention would make it possible to achieve them. Example 11: Evaluation of the antioxidant effect of a mixture of extracts of Vitis vinifera and a licorice extract against UVB.

[0138] The objective of this test is to compare the production of reactive oxygen species (ROS) by keratinocytes following chronic exposure to UVB in the presence of a composition according to the invention with that of a control.

[0139] To do this, fresh human skin fragments were immediately cut into 5 x 5 mm pieces and treated with trypsin-EDTA for 3 h at 37°C or overnight at 4C to separate the epidermis from the dermis. Keratinocytes were seeded at a concentration of 1x10 5 cells per cm 2 in 75cm 2 flasks in KGM-2 medium, which includes hydrocortisone (0.33 µg / ml), epidermal growth factor EGF (0.125 ng / ml), insulin (5 µg / ml), bovine pituitary extract (4 µl / ml), epinephrine (0.39 µg / ml), and transferrin (10 µg / ml). When cultures reached 70-80% confluence, cells were detached with 10% trypsin-EDTA, then resuspended in supplemented KGM-2 medium and counted on Malassez cells.The keratinocytes were then cultured at a rate of 5x10 4< cells per well in a 12-well plate for the cytoplasmic ROS production measurement test via a chloromethyl derivative of H2DCFDA, CM-H2DCFDA. Three days before UVB irradiation, a mixture comprising a composition according to the invention was placed in the presence or absence of the cells. The tested mixture was as follows: . 53.7% of a composition according to the invention itself composed of 53.4% ​​of E1, and 46.6% of E3 (the composition according to the invention providing 19.4% of flavanol monomers and 28 ppm of ε-viniferine) and 37.5% of a liquorice extract.

[0140] After changing the culture medium, the cells were irradiated or not a first time at 60mJ / cm 2 < , then new medium in the presence or not of the mixture of extracts was added and the cells were put back in the incubator at 37 ° C. Twenty-four hours after the first irradiation, the culture medium was changed and the cells were irradiated or not a second time at 60mJ / cm 2 < then new medium in the presence or not of the mixture of extracts was added and the cells were put back in the incubator at 37 ° C. Twenty-four hours after the second irradiation, the culture medium was changed and the cells were irradiated or not a third time at 60mJ / cm 2 < then new medium in the presence or not of the mixture of extracts was added and the cells were put back in the incubator at 37 C for 30 min.

[0141] Thirty minutes after the 3rd and final irradiation, cells were detached from the culture wells by trypsin-EDTA treatment, washed, and then incubated with 1µM / tube of CM-H2DCFDA for 30 minutes at 37°C in the dark. After this incubation, cells were washed twice in PBS and analyzed by flow cytometry. A minimum of 1x10 4 cells were collected and analyzed per assay. CM-H2DCFDA is used to indicate reactive oxygen species (ROS) in cells. It passively diffuses into cells, where its acetate groups are cleaved by intracellular esterases, and its thiol-reactive chloromethyl group reacts with intracellular glutathione and other thiols. The resulting oxidation produces a fluorescent adduct, which is trapped inside the cell.

[0142] The mean fluorescence intensity values ​​per cell were statistically compared using a two-way analysis of variance (ANOVA) where factor 1 was the treatment with the extract mixture or not, factor 2 was the irradiated treatment or not. A value of p < 0.05 is considered significant.

[0143] The results show that chronic irradiation at 60mJ / cm 2 < induces oxidative stress and significant ROS production. Indeed, the average fluorescence intensity per cell increases significantly from 1389 to 5346 after exposure to UVB. On the other hand, in the presence of a mixture of extract of Vitis vinifera and a licorice extract, no increase in the mean fluorescence intensity was observed, which means that there was no cytoplasmic production of ERO ( Figure 3 ).

[0144] These results demonstrate the antioxidant power and protective effect of a mixture of extract of Vitis viniferaand a licorice extract against UVB irradiation. Example 12 : Evaluation of cell viability following exposure to UVB and in the presence of a mixture of extracts of Vitis vinifera and a licorice extract.

[0145] The objective of this test is to compare the rate of cell proliferation of keratinocytes following chronic exposure to UVB in the presence of a composition according to the invention with that of a control.

[0146] After isolation of the keratinocytes, they are cultured at a rate of 5x10 3< cells per well in a 96-well plate. Three days before UVB irradiation, a mixture comprising a composition according to the invention was placed in the presence or absence of the cells. The mixture tested was as follows: 53.7% of a composition according to the invention itself composed of 53.4% ​​of E1, and 46.6% of E3 (the composition according to the invention providing 19.4% of flavanol monomers and 28 ppm of ε-viniferine), and 37.5% of a liquorice extract.

[0147] After changing the culture medium, the cells were irradiated or not a first time at 60mJ / cm 2 < , then new medium in the presence or not of the extracts was added and the cells were put back in the incubator at 37°C. Twenty-four hours after the first irradiation, the culture medium was changed and the cells were irradiated or not a second time at 60mJ / cm2 then new medium in the presence or not of the extracts was added and the cells were put back in the incubator at 37°C. Twenty-four hours after the second irradiation, the culture medium was changed and the cells were irradiated or not a third time at 60mJ / cm 2 < then new medium in the presence or not of the extracts was added and the cells were put back in the incubator at 37°C.

[0148] Twenty-four hours after the 3rd and final irradiation, 20µl of MTS was added to the wells, then the cells were returned to the incubator at 37°C and under 5% CO2 for 3 hours. At the end of the 3 hours, the absorbance (OD) was read at 490 nm by a spectrophotometer. The OD values ​​are then expressed as a percentage relative to the non-irradiated control for each condition.

[0149] Cell viability percentages were statistically compared using two-way analysis of variance (ANOVA) where factor 1 was treatment with the extract mixture or not, factor 2 was irradiated treatment or not. A value of p < 0.05 is considered significant.

[0150] The results show that chronic irradiation at 60mJ / cm 2 < induces a significant decrease in the number of viable cells. Indeed, the survival rate after exposure to UVB is 41.5%.

[0151] On the other hand, in the presence of a mixture of extract of Vitis vinifera and a licorice extract, no significant decrease in the number of living cells was observed after exposure to UVB ( Figure 4 ). These results demonstrate the protective effect of a composition according to the invention on cell viability following UVB irradiation. Example 13 : Examples of compositions intended for humans in different forms

[0152] Examples of compositions according to the invention are presented in Table 6 below. Example 14 : Nutritional drink intended for humans

[0153] Example 14 is a 50 ml drink containing the equivalent of 300 mg of a mixture of extracts of Vitis vinifera. The blend may consist of 225mg of grape seed extract and 75mg of white grape pomace extract. The extracts may be microencapsulated. The recommended dosage is 1 to 2 shots per day. Example 15 : Nutrient-enriched oil for humans

[0154] Example 15 is a vegetable oil consisting of grape seed oil containing extracts of Vitis vinifera and carotenoids. A daily dose of 20ml contains 150mg of a mixture of extracts of Vitis vinifera and 20mg of zeaxanthin. Example 16 : High-protein powder preparation

[0155] Example 16 is a formulation for a food intended for special medical purposes or a protein-enriched food formulation. The composition is detailed in Table 7. The dosage may be, for example, a daily dose of said powdered preparation. Said dose may be diluted in a drink or a dish and represents approximately 12g. Example 17 : Yogurt

[0156] Example 17 is a yogurt formulation, to be consumed 2 to 3 times a day. The composition is detailed in Table 7. [Table 7] Composition according to the invention Example 16 Example 17 Serving (g) 100 125 Constituents Catechin+epicatechin (mg), or 175mg of B (table 3) 50 62.5 ε-viniferine (mg) or 175mg of B (table 3) 0.0231 0.0289 Others Protein (g) 83 10 Vitamin D (µg) 125 15 Vitamin E (mg) 167 20 Vitamin B9 (µg) 3333 400 Vitamin B12 (µg) 833 100 Vitamin C (mg) 500 60 Zinc (mg) 42 5 Example 18 : Dog kibble

[0157] Example 18 is a formulation in the form of kibble intended for a 10 kg dog fed with 200 g of kibble / day providing: 0.450 mg of catechin and / or epicatechin per kg of body weight, and 0.045 µg of ε-viniferine per kg of body weight.

Claims

1. A composition comprising a mixture of molecules, comprising: - at least 15% of flavanol monomers, the percentage being given by dry weight relative to the total dry weight of the composition, and - at least 15 ppm of ε-viniferin, by dry weight relative to the total dry weight of the composition, said mixture of molecules is obtained from at least one extract or a mixture of extracts of Vitis vinifera, the ratio of flavanol monomers to ε-viniferin is between 20,000 and 4.

2. The composition according to the preceding claim, said composition comprising at least 100 ppm of stilbenes comprising said at least 15 ppm of ε-viniferin.

3. The composition according to either of the preceding claims, characterized in that the amount by weight of flavanol monomers is greater than the amount by weight of ε-viniferin.

4. The composition according to any of the preceding claims, characterized in that it comprises flavanol dimers (PAC B1 and B2) and in that the concentration of flavanol dimers (PAC B1 and B2) is greater than 5%, the percentage being given by dry weight relative to the total dry weight of the composition.

5. The composition according to any of the preceding claims, characterized in that the mixture of molecules is obtained from one or more natural product(s) selected from a Vitis extract, a tea extract, an apple extract, a cocoa extract, an iris, Sophora, Gnetum, Carex, peony, Dipterocarpus extract and a microalgae extract.

6. The composition according to the preceding claim, characterized in that the Vitis vinifera extract comprises at least 50% total polyphenols, the percentage being given by dry weight relative to the total weight of the Vitis vinifera extract.

7. The composition according to any of the preceding claims, characterized in that the mixture of molecules is obtained at least from one grape pomace extract and / or from a grape seed extract.

8. The composition according to the preceding claim, characterized in that the grape pomace extract(s) comprise flavonols and in that the concentration of flavonols in the composition is greater than or equal to 0.05% by dry weight relative to the total dry weight of the composition.

9. The composition according to any of the preceding claims, characterized in that it also comprises a licorice extract.

10. The composition according to any of the preceding claims, characterized in that the composition and / or one or more molecules of the composition is (are) encapsulated or microencapsulated in at least one food support selected from a maltodextrin, a gum arabic, a hydrogenated oil, a non-hydrogenated oil, a wax, an alginate, starch, a protein and mixtures thereof.

11. The composition according to any of the preceding claims, characterized in that it is in the form of a powder, gelcap, tablet, capsule, solution, suspension, emulsion, chewing gum, dairy product, cereals, cereal product or beverage.

12. The composition according to any of claims 1 to 11, for use thereof as a drug for humans or animals.