FR2935096A1 - Composition of fruit extract of Rosaceae family seeds, useful to prepare e.g. food composition, comprises polyphenols, chlorogenic acid and combination of flavonols (quercetin), dihydrochalcone (phloridzin), and flavanols (catechins) - Google Patents

Abstract

Composition (I) of fruit extract of Rosaceae family seeds comprises: polyphenols (at least 25 wt.% related to the total weight of dried material of the composition); chlorogenic acid (at least 15 wt.%); and a combination of at least 15 wt.% of flavonols (quercetin), at least 10 wt.% of dihydrochalcone (phloridzin), and at least 10 wt.% flavanols (catechins). Independent claims are included for a process of preparing (I) comprising: either (a) contacting the starting material dissolved in a solvent on a suitable adsorbent resin of styrene-divinylbenzene, (b) eluting a first fraction rich in hydroxycinnamic acids with an aqueous solvent comprising 15-25 wt.% of ethanol, and recovering the first fraction and then obtaining a powder of the first fraction by removal of aqueous solvent, (c) eluting a second fraction rich in flavanols with aqueous solvent comprising 35-45 wt.% of ethanol, recovering the second fraction and then obtaining a powder of the second fraction by elimination of aqueous solvent, (d) eluting a third fraction rich in dihydrochalcones and flavanols with aqueous solvent comprising 60-80 wt.% of ethanol, and recovering the third fraction and then obtaining a powder of the third fraction by elimination of the aqueous solvent, and (e) preparation of (I) by mixing the obtained powders in appropriate proportions; or (aa) preparation of a fraction rich in hydroxycinnamic acid, comprising (a1) contacting a first material dissolved in a solvent and adjusting to an acidic pH on the adsorbent resin of styrene-divinylbenzene, and again adjusting to an acidic pH, where the acidic pH is 2-3, (a2) elution of first fraction rich in hydroxycinnamic acid with an aqueous solvent comprising ethanol (30-50%), recovering the first fraction, and obtaining a powder of first fraction by elimination of aqueous solvent, (bb) preparation of second fraction rich in flavonols and third fraction rich in flavonols and dihydrochalcones comprising (b1) contacting a first material dissolved in a solution in a appropriate solvent and adjusting to an basic pH on the adsorbent resin of styrene-divinylbenzene and again adjusting to an basic pH, where the basic pH is 6.5-7.5, (b2) elution of second fraction rich in flavonol with an aqueous solvent comprising ethanol (35-45 wt.%), recovering the second fraction, and obtaining a powder of second fraction by elimination of aqueous solvent, (b3) elution of a third fraction rich in flavonol and dihydrochalcones with an aqueous solvent comprising ethanol (65-75 wt.%), recovering the third fraction, and obtaining a powder of third fraction by elimination of aqueous solvent, and (cc) preparation of (I) by mixing the obtained powders in appropriate proportions. ACTIVITY : Nootropic; Dermatological; Osteopathic; Hypotensive; Anorectic; Antiarteriosclerotic; Anticoagulant. MECHANISM OF ACTION : None given.

Description

FIELD OF THE INVENTION The present invention relates to the field of the manufacture of compositions containing polyphenols, intended for food, dietary, nutraceutical and cosmetic use.

PRIOR ART Polyphenols have long been known for their beneficial activities in many fields, including the medical, cosmetic and food fields. In general, polyphenols constitute a broad family of molecules having in common the presence in their more or less complex structure of a plurality of phenolic groups. Polyphenols include various molecular classes including phenolic acids, flavanones, tannins, and anthocyanins. Many plant extracts enriched in polyphenols have been described in the state of the art as well as their methods of preparation. The most representative are the extracts of grape pomace, grape skins, grape seeds, tea, pine bark, olive and rosemary (see patent documents FR 2 643 073, FR 2 372 823, US 4,698,360, EP 348,781, EP 283,349, FR 1,427,100, FR 2 092 743). Each class of polyphenols has an activity, or a combination of activities, on the physiology of an organism that is specific and at least substantially distinct from the activities of other classes. For example, extracts rich in chlorogenic acid, which compound is found in large quantities in apple, coffee or green tea, are known to inhibit the intestinal assimilation of sugar and their inhibitory effects. on weight gain. Phloridzin enriched extracts have been previously described for their effects against diabetes. However, recent studies with highly purified phloridzine show that this polyphenol is not active in vivo in humans. Quercetin-enriched extracts, including some quercetin glycosides, have been described for their effects on glucose metabolism, and for their antihypertensive effects.

Extracts rich in catechins have been described for their effects against cancers or against inflammatory or coronary diseases. However, it is observed that the in vivo effects of each class of polyphenols which have been described in the state of the art have been obtained after administration of extracts enriched in certain polyphenols and not after administration of a given polyphenol in purified form. Plant extracts enriched in polyphenols thus comprise various combinations of polyphenols, both qualitatively and quantitatively. Thus, plant extracts enriched in polyphenols include one or more quantitatively major polyphenols, in combination with a plurality of other minority polyphenols. Because of the variable nature of the qualitative and quantitative polyphenol composition of these plant extracts, the actual contribution of a particular polyphenol type to the effect that is observed in vitro or in vivo is difficult, and in some cases impossible, to determine. Indeed, the existence of in vitro or in vivo synergistic effects between a quantitatively predominant polyphenol and one or more quantitatively minor polyphenols can not be ruled out. In general, depending on the type of quantitatively major polyphenol desired in a final plant extract, said extract is prepared from plants which are known to naturally contain a high content of the polyphenol type of interest. For example, some pome fruits of the family Rosaceae, such as the apple, are known to be naturally rich in flavanols, and more particularly rich in catechins and oligomers and catechin polymers, and to a lesser extent also rich in hydroxycinnamic acids including chlorogenic acid. The apple is also known to have a low content of dihydrochalcones, including phloridzine, especially in the edible part of the fruit. However, given the complex qualitative and quantitative constitution of apple polyphenols, the contribution of each type of polyphenol to its overall effects in vitro or in vivo, for example the effects on glycemia, does not seem to be determined today. . Generally speaking, the plant extracts, including apple extracts, are generally found in the quantitative ratios of a polyphenol to another polyphenol which are initially present in the fruit. For example, the polyphenolic apple extract described in patent document EP 657 169, which essentially contains the water-soluble compounds which pass easily in the juice, namely those of the group of hydroxycinnamic acids (chlorogenic, caffeic, p-coumaric). ) and the flavanol group (catechins and catechin polymers, also called procyanidols). Quercetin and the glycosylated derivatives of phloretin (phloridzine) are present in small proportions. In specific cases, it is sought to enrich the final extract composition, selectively into one or more types of polyphenols. There may be mentioned, in particular, the patent document FR 2 822 466 which describes a process for obtaining polyphenols from apple as well as the final product extract of the process, which is enriched in phloridzine. PCT document WO 2007/026101, which describes methods for obtaining polyphenolic extracts from apple, and more precisely is precisely from various constituents of the apple such as the skin, the pulp or the seeds, allowing the preparation of final compositions of qualitative and quantitative constitutions varied in polyphenols. This document describes, for example, an extract obtained from apple pulp which is enriched in flavanols and which also contains hydroxycinnamic acids, dihydrochalcones and flavonols. This extract of apple pulp comprises a maximum content of catechins and catechin polymers of 10% by weight of the total polyphenols and a maximum content of dihydrochalcones of 10% by weight of the total polyphenols. An extract enriched with phloridzin obtained from whole apple seeds is also described. Also disclosed is a flavonol-enriched extract and a wax-enriched extract obtained from apple peels. There is still a need in the state of the art for new compositions containing polyphenols, especially for use in the medical, cosmetic and food. SUMMARY OF THE INVENTION The present invention relates to a pome fruit extract composition of the Rosaceae family comprising at least 25 percent by weight of polyphenols based on the total dry weight of said composition, said composition having a content of at least 15% by weight of hydroxycinnamic acids and said composition comprising a combination of: - at least 15% by weight of flavonols (quercetin), - at least 10% by weight of dihydrochalcones ( phloridzine), and - at least 15 percent by weight of flavanols (catechins), the weight percentages of the hydroxycinnamic acids, flavonols, dihydrochalcones and flavanols being expressed relative to the total weight of the polyphenols. This invention also relates to the use of the above extract composition for the manufacture of pharmaceutical, cosmetic, food, cosmeceutical or even nutraceutical compositions. The present invention also relates to processes for the manufacture of a pome fruit extract composition of the Rosaceae family as described above. DETAILED DESCRIPTION OF THE INVENTION The Applicant has sought to define the optimal qualitative and quantitative constitution of polyphenols for a composition capable of exerting a combination of health benefits. Specifically, the applicant has carried out important research to develop compositions of polyphenolic extracts obtained from pome fruits belonging to the family Rosaceae, and particularly from apples, and whose qualitative constitution. and specific quantitative polyphenols allows to obtain a combination of beneficial effects for human or animal health, including the following effects: anti-oxidant effects, regulation of blood glucose and regulation of lipid metabolism. According to the invention there is provided a composition comprising an apple extract enriched in polyphenols and having a minimum and balanced content of certain polyphenols, said composition having, in particular, advantageous activities on glycemia and lipid metabolism. The subject of the invention is a composition of pome fruit extract of the family Rosaceae comprising at least 25 percent by weight of polyphenols relative to the total weight of dry matter of said composition, said composition having a content of at least 15 percent by weight of hydroxycinnamic acids and said composition comprising a combination of: - at least 15 percent by weight of flavonols (quercetin), - at least 10 percent by weight of dihydrochalcones (phloridzin), and - at least 10 weight percent flavanols (catechins), the weight percentages of hydroxycinnamic acids, flavonols, dihydrochalcones and flavanols being expressed based on the total weight of the polyphenols. In the present description, the overall contents of polyphenols are determined according to the conventional Folin Ciocalteu technique. In the present description, the contents of each of the polyphenols of dihydrochalcone, flavonol, flavanol and hydroxycinnamic acid types are determined by the conventional technique of high pressure liquid chromatography (HPLC). In the present description, the levels of hydroxycinnamic acids are expressed in chlorogenic acid equivalent. In the present description, the flavonol contents are expressed in quercetin equivalent. In the present description, the dihydrochalcone contents are expressed as phloridzin equivalent. In the present description, the flavanol contents are expressed in catechin equivalents. Surprisingly, it has been shown according to the invention that the pome fruit extract composition of the Rosaceae family above, when administered in vivo, has beneficial effects on blood glucose. In particular, it has been shown in the examples that the pome fruit extract composition of the Rosaceae family above is capable of regulating the postprandial glucose level, and more precisely the last phase of postprandial glucose, this is that is, about one hour to two hours after the meal. In addition, it has been shown that the above-mentioned Rosaceae family of pome fruit extract has reduced cytotoxicity, in comparison with the cytotoxicity of other polyphenolic apple extracts, particularly polyphenolic extracts having unbalanced in certain polyphenols, including a polyphenolic apple extract enriched in phloridzine. It has also been shown according to the invention that the pome fruit extract composition of the Rosaceae family above has cell differentiation inhibiting activities, particularly an activity of inhibiting adipocyte differentiation. It has also been shown that the pome fruit extract composition of the Rosaceae family above has other activities related to lipid metabolism, including lipogenesis inhibiting activity and lipophilic activity. inhibition of lipid accumulation. In particular, it has been shown that the activities of the pome fruit extract composition of the Rosaceae family above in relation to lipid metabolism is not correlated with the dihydrochacone content, and in particular with the phloridzine content. Without wishing to be bound by any theory, the plaintiff thinks that, in the overall effect of the pome fruit extract composition of the Rosaceae family above, which is brought about by the combination of the different types of polyphenols Since this composition contains, the minimum content of hydroxycinnamic acids, in particular chlorogenic acid, and the minimum content of flavonols, in particular quercetin, have an important role. Moreover, the pome fruit extract composition of the Rosaceae family above has an original constitution in that it comprises a high minimum content of both hydroxycinnamic acids, including chlorogenic acid, and dihydrochalcones. , including phloridzin, and flavonols, including quercetin. It has also been shown in the examples that the pome fruit extract composition of the Rosaceae family above has no proinflammatory effect. Therefore, because of its reduced cytotoxicity and the lack of a pro-inflammatory effect, the above-mentioned Rosaceae seed pome fruit extract composition is likely to be administered safely to humans. human or animal, especially non-human mammals, since few adverse effects or even no adverse effects are likely to be caused.

It has also been shown according to the invention that the pome fruit extract composition of the Rosaceae family above causes little or no fluctuations in the expression of the genes encoding the endothelial (eNOS) and neuronal or central isoforms, respectively. (cNOS) of the nitric oxide synthetase enzyme, confirming that said composition is not likely to cause adverse effects. In addition, the low and transient overexpression of the eNOS isoform that is induced by the above-mentioned Rosaceae seed pome fruit composition would be in favor of a potential beneficial effect in the case of hypertension. as well as a potential beneficial anti-thrombotic and anti-atherosclerotic effect. In addition, the combined antioxidant effects of the various polyphenols included in the pome fruit extract composition of the Rosaceae family above are capable of imparting to said composition an important value for reducing the content of the organism by free radicals, including intracellular free radicals, which can be generated in pathological or non-pathological situations, including during sports training. Thus, the extract composition of the invention is capable of promoting a beneficial effect on carbohydrate metabolism, glyconeogenesis, thermogenesis and respiratory functions. Without wishing to be bound by any theory, the Applicant believes that the various beneficial activities of the Rosaceae seed extract composition above could be related in particular to the minimal hydroxycinnamic acid content, which is significantly 25 greater than the content of these compounds which is conventionally found in the known polyphenolic apple extracts. In addition, the Applicant believes that the various beneficial effects of the Rosaceae seed pome fruit extract composition above are likely to be related to the balanced amounts of each of the four major types of polyphenols that such composition. contains, which, to the knowledge of the applicant, are not found in any other polyphenol extract of apple having these minimum levels of polyphenols. In other words, the Applicant believes that the various beneficial effects of the above Rosaceae family of pome fruit extract composition are due both to (i) the balanced nature of the hydroxycinnamic acid contents, flavonols, dihydrochalcones and flavanols and (ii) the minimum content of each of these four types of polyphenols, which, without causing toxicity to the individual, is physiologically active. It is also noted that the composition of pome fruit extract of the Rosaceae family above includes a minimum content of hydroxycinnamic acids, including chlorogenic acid, polyphenol known for its activity of regulating the metabolism of sugars, and especially for its activity of inhibition of glyconeogenesis. It is also possible to emphasize the minimum content of flavonols, including quercetin, polyphenols known for their glycemic inhibition activity. According to some preferred embodiments, the pome fruit extract composition of the Rosaceae family above comprises at least 20, 25, 30, 35, 40, 45, 50 or 55 percent by weight of hydroxycinnamic acids. , relative to the total weight of the polyphenols contained in said composition. In certain embodiments, said composition may comprise 60 percent by weight of hydroxycinnamic acids, based on the total weight of the polyphenols. According to some preferred embodiments, the above-mentioned Rosaceae seed fruit extract composition comprises at least 15, 20, 20 or 30 percent by weight of flavonols, including quercetin, relative to total weight of polyphenols. In some embodiments, said composition may comprise 35 percent by weight of flavonols, including quercetin, based on the total weight of the polyphenols. In some embodiments, the pome fruit extract composition of the Rosaceae family above comprises at least 15 or 20 percent by weight of dihydrochalcones, including phloridzin, based on the total weight of the polyphenols. In some embodiments, said composition may comprise 25 percent by weight of dihydrochalcones, including phloridzin, based on the total weight of the polyphenols. According to some embodiments, the pome fruit extract composition of the Rosaceae family above comprises at least 15 or 20 percent by weight of flavanols, including catechins, based on the total weight of the polyphenols. In some embodiments, said composition may comprise 25 weight percent flavanols, including catechins, based on the total weight of the polyphenols.

In another aspect, the pome fruit extract composition of the Rosaceae family above comprises at least 30, 35, 40, 45, 50, 55, 60, 65, 70 or 75 percent by weight of polyphenols. total, relative to the total weight of dry matter of said composition. In some embodiments, said composition may comprise 80 percent by weight of polyphenols, based on the total dry weight of said composition. In some preferred embodiments, the pome fruit extract composition of the Rosaceae family above comprises at least 25 percent by weight of total polyphenols, for example from 30 to 80 percent by weight of total polyphenols. relative to the total weight of dry matter of the composition, and comprises: - at least 15 percent by weight, for example from 30 to 60 percent by weight, of hydroxycinnamic acids (chlorogenic acid), - at least 15 percent by weight; percent by weight, for example from 25 to 35 percent by weight, of flavonols (quercetin), - at least 10 percent by weight, for example from 20 to 30 percent by weight, of dihydrochalcones (phloridzin and other phloretin), and - at least 10 percent by weight, for example from 15 to 25 percent by weight, of flavanols (catechins), the weight percentages of hydroxycinnamic acids, flavonols, dihydrochalcones and flavanols being expressed in relation to the total weight of polyphenols. In some preferred embodiments, the above-described Rosaceae seed fruit extract composition comprises a substantially equivalent amount of each of the four types of polyphenols of interest. Thus, the preferred extract compositions of the invention include those compositions in which the polyphenols of interest are present in the following amounts: 20 to 30 percent by weight of hydroxycinnamic acids, including 30 of chlorogenic acid from 20 to 30 percent by weight of flavonols, including quercetin, from 20 to 30 percent by weight of dihydrochalcones, including phloridzin, and from 20 to 30 percent by weight of flavonols, including the percentages by weight being expressed relative to the total weight of said polyphenols of interest in the composition. A particularly preferred composition of the invention comprises a content of 25 percent by weight of each of the four polyphenols of interest, based on the total weight of said polyphenols of interest in the composition. Various methods have been developed according to the invention in order to directly obtain, from a raw material of pips fruit of the Rosaceae family, the extract compositions described above. Specific illustrations of these methods are described in the examples. The characteristics of the various processes for obtaining the extract compositions which are disclosed in the present description are not necessarily limiting. The methods which are disclosed in this specification, and particularly the methods described in the examples, may be adapted by those skilled in the art in view of his general knowledge. The pome fruits of the Rosaceae family are preferably chosen from apples, quinces, loquats, pears and corms. Preferably, an extract composition according to the invention consists of an apple extract composition, and most preferably apple cider. In general, an extraction process for obtaining the extract compositions according to the invention is characterized by the fact that mature or non-mature apples, by-products and by-products of apple processing are used. , different apple tissues (skins, seeds, pulp). Preferably, cider apples are used, and particularly cider apples chosen from the varieties currently exploited in western France, mainly in Normandy and Brittany. As an illustration, cider apples of a variety chosen from the following varieties are used: Bitter New, Armagnac (Sour), Sweet Corier, Marseigna, 30 Normandy White, Apple Orchard, Blanquette, Dello, Doudazin, Germaine, Mom Lili, Normandy White; Pass Queen, Peaud'Ane, Apple orchard, Pear, President or Roguet Rouge Judor, Little Yellow, Binet Red, Juliana, Clozette, Avrolles, Dous Moen, Antoinette, Bedan, Dabinett, Sweet Coet Lined, Yellow Knight, Jeanne Fox , Kermerrien.

Preferably, the pome fruit extract composition of the Rosaceae family above is in the form of a powder, for example a fine powder or a granulated powder. The present invention also relates to a process for obtaining a composition of pome fruit extract of the family Rosaceae as described above, said process comprising the following steps: a) fresh or dry raw materials are ground to the desired particle size. b) the ground product is pressed and / or extracted with water or a water / organic solvent mixture. The extraction solutions can be made acidic or basic by providing the appropriate reagent. The organic solvents may be chosen from ethanol, methanol, acetone, ethyl acetate, hexane and petroleum ether. Preferably, solvents compatible with human nutrition, such as agricultural ethanol, are used. C) the pressing or extraction residues from step b) can be further extracted with the same or a different solvent. d) the pressing and extraction juices obtained in step b) and in step c) when step c) is carried out, are combined, optionally acidified and then clarified by centrifugation, filtration, ultrafiltration or any other technique 20 for performing the operation. e) the pressing and extraction juices may undergo a gluing step to specifically remove most of the catechin polymers (tannins). f) the clarified juice obtained in step d), and / or in step e) when step e) is carried out, is optionally distilled to remove the extraction solvent and then concentrated and dried. g) In order to improve the level of polyphenols in the final powder, the juice freed from any organic solvents obtained in step f) is preferably passed over a support type adsorbent resin, ion exchange resin, polyvinylpropylene (PVP), activated charcoal or any other support for selective adsorption / desorption of polyphenols. If necessary, the equilibration pHs of the adsorbent and clarified juice can be adapted for more selective adsorption / desorption of the different groups of polyphenols. g) the adsorbent / polyphenol support complex is washed with acidulated water or not to eliminate undesirable compounds (sugars, polysaccharides, amino acids, organic acids, proteins, minerals, etc.). h) the acidulated water and residual contaminants retained in the dead volume of the adsorbent support are removed by washing with water. i) the polyphenols are desorbed from the adsorbent support by an ethanol solution or any organic solvent such as acetone, methanol, ethyl acetate or acid / base solutions to achieve this objective. j) for selective separation of certain polyphenol groups preferentially use successive stages of increasing concentrations of solvent. k) the liquid phases obtained, containing the polyphenols, are distilled to separately recover the solvents and polyphenolic concentrates. I) the concentrates obtained are spray dried, lyophilized or any other drying process to obtain a powder preserving the quality of the assets. In some of its embodiments, the method for obtaining a pome fruit extract composition of the Rosaceae family comprises the following steps: a) extraction with water, with the water / solvent mixture or to the organic solvent b) distillation of the extract to remove the organic solvent c) pH adjustment, clarification and / or bonding of the desolventized extract d) purification and / or fractionation of the adsorbent supported polyphenols allowing them total or selective adsorption / desorption; e) distillation and concentration making it possible to eliminate the organic desorption solvent; f) mixing the concentrates of different polyphenolic groups according to the desired proportions. (g) appropriate drying preserving the quality of the polyphenols. With the above process, optimal extraction and solubilization conditions of the polyphenolic compounds have made possible the production of the extract compositions defined in this specification. A particularly interesting feature of some embodiments of the method described above is the fact that the principle of step chromatography is used to separate the polyphenol groups and thus largely exclude certain compounds such as procyanidins while preserving the other components in the final product. As shown in the examples, and in particular in Examples 5 and 6 below, extract compositions according to the invention can be obtained by (i) selective extraction of the various types of polyphenols of interest, then (ii) mixing the selective extraction products in selected proportions. The subject of the present invention is also a method for the manufacture of a composition of pome fruit extract of the Rosaceae family comprising the following steps: a) contacting the starting raw material in solution in a suitable solvent on a styrene-divinylbenzene adsorbent resin; b) eluting a first fraction enriched in hydroxycinnamic acids with an aqueous solvent comprising from 15% to 25% by weight of ethanol, and 1s recovering said first fraction, and then obtaining a powder of said first fraction by eliminating the aqueous solvent, c) eluting a second fraction enriched in flavanols with an aqueous solvent comprising from 35% to 45% by weight of ethanol, and recovering said second fraction, and then obtaining a powder of said second fraction by elimination of the aqueous solvent, d) elution of a third fraction enriched in dihydrochalcones and flavonols with an aqueous solvent comprising from 60% to 80% by weight of ethanol, and recovery of said third fraction, then obtaining a powder of said third fraction by removal of the aqueous solvent, e) preparation of a pome fruit extract composition of the family Rosaceae as defined in the present description by mixture of the powders obtained respectively in steps b), c) and d), in appropriate proportions. The starting raw material may be selected from apple pomace, apple-skin enriched fraction, whole apple pips, seed shell, peeled kernel kernels, apple juice, or a mixture of two. , or more than two, of the abovementioned raw materials. Preferentially, the starting raw material consists of an apple juice, for example a raw apple juice, or an apple juice in which the suspended solids have previously been removed. The suspended solids may be removed by conventional techniques such as clarification or gluing. Advantageously, the adsorbent resin used in step a) insists on a styrene-divinylbenzene type resin having an average pore diameter of 100 Angstroms. By way of illustration, it is possible to use the Amberlite XAD-1600N resin marketed by Rohm and Haas (Paris, France). Preferably, the solvent used in step a) is water. In the embodiments of the process in which the starting raw material is an apple juice, the water solvent is already contained in said starting raw material. Preferably, the raw material in liquid suspension or in solution is adjusted to a pH ranging from 6 to 7.5, preferably to pH 7. Preferably, the solvent used in steps b), c) and d), in the solvent mixture / ethanol, is water.

In step b), the elution is preferably carried out with a solvent comprising 20% by weight of ethanol. In step c), the elution is preferably carried out with a solvent comprising 40% by weight of ethanol. In step d), the elution is preferably carried out with a solvent comprising 70% by weight of ethanol. In each of steps b), c) and d), the removal of the solvent is preferably carried out by distillation and then optionally final drying, for example by drying in an atomization tower. Advantageously, each of steps b), c), d) and e) is followed by a step of analyzing the content of the powder obtained with polyphenols, and in all cases the content in each of the polyphenol classes of polyphenols. interest, namely hydroxycinnamic acids, flavonols, dihydrochalcones and flavanols. In step e), a pome fruit extract composition of the Rosaceae family as defined in the present description, preferably an apple extract composition, is prepared by adjusting the amounts of each of the powders. the first, second and third fractions to have the desired content of polyphenols in the final composition. At the end of step e), it is possible to obtain, if desired, an extract composition according to the invention comprising 25% by weight of each class of hydroxycinnamic acid polyphenols, flavonols, dihydrochalcones and flavanols, relative to the weight total of the above polyphenols. The present invention also relates to a process for the manufacture of a pome fruit extract composition of the family Rosaceae comprising the following steps: a) preparation of a fraction enriched in hydroxycinnamic acids according to a process comprising the steps following: a1) contacting the starting raw material in solution in an appropriate solvent and adjusted to an acidic pH on a styrene-divinylbenzene adsorbent resin also adjusted to an acid pH, the acidic pH being a pH ranging from from 2.0 to 3.0, preferably 2.5, a2) eluting a first fraction enriched in hydroxycinnamic acids with an aqueous solvent comprising from 30% to 50% by weight of ethanol, and recovering said first fraction , then obtaining a powder of said first fraction by removal of the aqueous solvent, b) preparation of a second fraction enriched in flavanols and a third fraction enriched in f lavonols and dihydrochalcones, according to a process comprising the following steps: b1) contacting the starting raw material in solution in a suitable solvent and adjusted to a basic pH on a styrene-divinylbenzene type adsorbent resin also adjusted to pH basic, the basic pH consisting of a pH ranging from 6.5 to 7.5, b2) eluting a second fraction enriched in flavanols, with an aqueous solvent comprising from 35% to 45% by weight of ethanol, and recovery of said second fraction, then obtaining a powder of said second fraction by removal of the aqueous solvent, b3) eluting a third fraction enriched in flavonols and dihydrochalcones, with an aqueous solvent comprising from 65% to 75% by weight of ethanol, and recovering said third fraction, then obtaining a powder of said third fraction by removal of the aqueous solvent, c) preparing a pome fruit extract composition of the Rosaceae family as defined in the present description by mixing the powders obtained respectively in steps a) and b), in appropriate proportions.

The general conditions of the above process, in particular the nature of the starting raw materials, the nature of the solvents used, the nature of the adsorbent resin, etc., are the same as those described for the process for obtaining enriched fractions. in polyphenols which has been previously described.

In step a1) the acidic pH is advantageously 2.5. In step a2), use is preferably made of an aqueous solvent comprising 40% by weight of ethanol. In step b1), the basic pH is preferably 7. In step b2), use is preferably made of an aqueous solvent comprising 40% by weight of ethanol. In step b3), use is preferably made of an aqueous solvent comprising 70% by weight of ethanol. In step c), a composition of pome fruit extract of the Rosaceae family as defined in the present description is prepared, preferably a composition of apple extract, by adapting the amounts of each of the powders of the first, second and third fractions to have the desired content of polyphenols in the final composition. At the end of step c), it is possible to obtain, if desired, an extract composition according to the invention comprising 25% by weight of each class of hydroxycinnamic acid polyphenols, flavonols, dihydrochalcones and flavanols, relative to total weight of the above polyphenols. The extracts according to the invention are obtained by the process described above, preferably from apples or other fruits of the family Rosaceae, their by-products, specific fractions of tissues (hides, pulp, pips, 25 seed shells) or a suitable mixture of these fractions. The purpose of this mixture is to obtain a rich and balanced extract in the three desired components that are chlorogenic acid, quercetin and phloridzin. Food grade plant extracts having the characteristics described and obtained according to the process described in the present invention are useful in oral or topical applications as food ingredients, food supplements, nutraceuticals or cosmetics. The extracts rich in chlorogenic acid and balanced in quercetin and phloridzin according to the invention have antioxidant, free radical-free, anti-UV properties. More particularly, the extracts according to the invention exhibit regulating properties of intestinal and cellular assimilation of nutrients such as sugars. These extracts also regulate liver glyconeogenesis and generally exhibit weight control properties, slimming, anti-cellulite and anti-obesity effects. By their composition and their properties, the extracts according to the invention can also be used in formulations for respiratory comfort and the management of physical effort. The present invention also relates to food compositions, dietary supplements, nutraceuticals, cosmetics comprising, among others, one of the extracts rich in chlorogenic acid and equilibrated with quercetin and phloridazine as previously described. An interesting originality of the extracts according to the present invention is that they are particularly depleted in catechin polymeric procyanidins. As a result, the undesirable effects of these compounds in food and cosmetic formulations, namely color, bitterness, astringency, the tanning and complexing power of proteins, are substantially reduced. The compositions according to the invention can be used orally or topically. Depending on the mode of use, the compositions according to the invention will be in one of the forms usually used in oral ingestion or in cosmetics. The compositions according to the invention may especially be formulated in the form of capsules, capsules, lozenges, milks, lotions, creams, gels, beverages, dairy products, confectionery, biscuits or any other formulation allowing their oral or topical use. The food, dietary supplement, nutraceutical or cosmetic compositions of the present invention are formulated by the methods and technologies traditionally used by those skilled in the art depending on the applications for which they are intended.

Characteristics of the compositions comprising a pome fruit extract composition of the Rosaceae family In the case of dermatological applications, the compositions according to the invention can be combined with other known compositions, such as sunscreen, to effectively manage the condition of the skin. The dermatological compositions produced according to the present invention comprise at least one of the apple extracts obtained according to the process of the invention, or else apple extracts obtained according to the teaching of PCT Application No. WO 2007/026101. at a concentration sufficient to obtain the desired effects. For extracts rich in polyphenols and / or ursolic acid, it will be necessary to provide sufficient amounts to neutralize free radicals and observe an anti-oxidative, anti-aging, anti-aging effect. For polyphenolic extracts particularly rich in chlorogenic acid, quercetin and / or phloridzin, in addition to the action against free radicals, the compositions can provide sufficient amounts to obtain beneficial effects in the regulation of lipid metabolism and carbohydrate, thinness, anti-cellulite action and anti-osteoporosis action. In addition to the apple active ingredients, the compositions according to the present invention may comprise carriers / vectors, texturizing agents, colorants, flavorings, sanitary and pharmaceutically acceptable preservatives, known to those skilled in the art. In the case of nutraceutical, food, food supplement, and cosmeceutical applications, the compositions according to the present invention are used alone or in combination with other beneficial agents, at concentrations sufficient to obtain the desired effects against, for information purposes only. body aging condition, osteoporosis, obesity, excess weight. By way of indication but without limitation, the desired effects may be the neutralization of free radicals, the restoration or increase of the (potential) plasma antioxidant status, the reduction of oxidative stress, the improvement of tissue remodeling, particularly dermal remodeling. , weight reduction, prevention of osteoporosis and adverse effects of menopause, prevention or reduction of cellulite. In the topical and / or oral compositions, according to the present invention, the apple extract (s) are incorporated at physiologically effective levels of between 0.01 and 50%, preferably between 0.1 and 10%, more preferably between 0 and 10%. , 2 and 5%. Any vector and / or formulation carrier, texturizer, colorant, stabilizer, flavor, preservative, in accordance with the use and regulation of the products intended for consumption or topical application can be used in the production of the compositions. The topical compositions according to the present invention may comprise one or more moisturizing agents commonly used to facilitate hydration of the skin and to inhibit or prevent skin loss in water. The topical composition may also include exfoliating acid components usually used by those skilled in the art such as fruit acids.

The term physiologically effective rate means here the amount of apple active to obtain the expected benefits in the management of the dermatological state or the general condition of the body. It is understood by those skilled in the art that this amount of active ingredients may vary depending on the individual and his skin condition. The importance of the doses of active ingredients or the number of daily uses of a given composition will depend on the sensitivity of the persons and the route of administration. Doses and their frequency may also be a function of the age, weight, health status and response of each individual. For oral administration, the incorporation rates of the apple active ingredients will be between 5 mg and 1000 mg, preferably between 25 and 500 mg and more preferably between 50 and 400 mg. The routes of administration of apple actives are those generally recommended by those skilled in the art. Preferably, the topical and oral routes are used. The forms of presentation of the compositions may be, by way of indication but without limitation, tablets, capsules, capsules, ampoules, dispersions, suspensions, aerosol solutions, spray, sponges, applicator cottons, patches, pastes, gels, lotions, syrups, ointments, powders, granules, creams. The compositions according to the invention may be prepared by either of the methods known to those skilled in the art. These methods include the combination of a carrier / vector with apple actives, as well as other ingredients, additives and processing aids necessary to achieve stable formulas adapted to the intended mode of use. The active ingredients and other ingredients are formulated by the apparatus and in the usual orders of blends, agitation, texturing, emulsification, expansion, packaging, etc. known to those skilled in the art. In addition to the usual forms of formulation and conditioning, the compositions according to the invention can be produced in particular formulations adapted for the protection of the active ingredients, their transport in the body and their gradual release and controlled over time at the level of the target area, as described for example in US Patents 6,630,163; US 5,591,767; US 3,536,809; US 4,008,719; US 5,354,556. The particular forms of formulation may comprise a step of encapsulation, entrapment in another macromolecule or a structure of molecular complexes of the cyclodextrin, liposome, lipoprotein globule or gelled micro-bead types. The particular forms of stabilized formulation may also include a covalent bonding step between the active ingredients of the apple extract and another protective or transport molecule such as a fatty acid ester or proteinaceous linkage.

The examples below illustrate the processes, extracts and applications of the invention without being limiting.

EXAMPLES Example 1: Preparation of an apple extract composition A volume of 100 kg of dry pomace of apple was extracted with 1400 liters of water heated at 70 ° C. for 1 hour. The mixture is filtered off through a 100 μm sieve to obtain 1100 liters of filtrate. The filtrate is clarified on a plate centrifuge to obtain 1078 liters. This clarified volume is microfiltered on an ultrafiltration pilot plant is equipped with tubular mineral membranes to recover 1000 liters of permeat. The permeate is passed over a column of 15 liters of adsorbent resin XAD-1600 previously washed and equilibrated in water. The discolored eluate is collected. After passing all the ultrafiltration permeate, the column is washed with 2 volumes of hot water. The discolored eluate and the wash water are mixed and acidified with 0.5% citric acid. This preparation contains essentially chlorogenic acid not fixed by the resin. Ethanol diluted to 25% is passed over the column to remove most of the catechins and their procyanidol polymers. The alcohol, the catechins and their residual polymers retained in the dead volume of the column are removed by washing with 2 volumes of water. The column is then equilibrated with 2 volumes of 0.5% citric acid solution in water. The discolored and acidified eluate from the passage of the permeate UF is ironed on the column to fix the phenolic acids (essentially chlorogenic acid). The undesirable elements (sugars, malic acid, minerals, amino acids, etc.) are removed by washing the column with 2 volumes of acidulated water.

The citric acid retained in the dead volume is removed by washing with 2 volumes of water. The polyphenols retained in the column are eluted by passing 1.5 volumes of a 70% diluted ethanol solution.

The ethanol and the residual polyphenols retained in the dead volume of the column are evacuated by shoot with 1.5 volumes of water. The entire alcoholic extract (45 liters) is distilled and concentrated on a rotary evaporator to obtain 1 kg of concentrate. The concentrate is frozen and lyophilized to recover 295 g of powder. The analysis of the polyphenols in the powder gives the following results: 33.4% of total polyphenols by Folin colorimetric assay with epicatechin as a standard or 69.5 with chlorogenic acid as a standard. 23.46% of total polyphenols by HPLC assay of which 15 ° 18.5% of hydroxycinnamic acids (chlorogenic acid) ° 21.6% of flavonols (quercetin) ° 16.8% of dihydrochalcones (phloridzin) ° 43.1% flavanols (catechins and catechin polymers)

Example 2: Preparation of apple extract composition Dried apple marc was fractionated to obtain 2100 Kg of a very skin enriched fraction. The skins are extracted for 1 hour with 27900 liters of water heated to 70 ° C. The solids are then separated from the liquid phase by passage over a horizontal decanter followed by a centrifuge clarifier to obtain 24,000 liters. The clarified phase is ultrafiltered on 10,000 Dalton organic spiral membranes to recover 22,800 liters of permeate. The pH of the permeate is adjusted to 7 with dilute sodium hydroxide and a 200 liter column of XAD adsorbent resin is also equilibrated to neutral pH. The neutralized permeate is passed over the column to selectively bind the basic or neutral polyphenols. The unfixed phenolic acids, more particularly the hydroxycinnamic acids, are collected in the eluate.

30% ethanol in water is passed over the column to remove most of the catechins and procyanidols. The column is then acidified and equilibrated with a dilute hydrochloric acid (HCI) solution.

The eluate containing the phenolic acids is also acidified to pH 2.5 with dilute hydrochloric acid. This acidified solution is ironed on the column to fix the hydroxycinnamic acids. The undesirable elements, especially the sugars, are removed by washing the column with acidulated water. All the polyphenols retained in the column is recovered by passing 1.5 volumes of 70% ethanol solution followed by 1.5 volumes of water. The whole of this ethanol extract (600 liters) is distilled and concentrated to obtain 42 kg of concentrate. This concentrate is spray dried. 5.6 kg of powder is thus recovered. The analysis of the polyphenols gives the following results: 36.7% of total polyphenols by colorimetric assay with Folin with epicatechin as standard or 47.8% with chlorogenic acid as standard. 26.7% of total polyphenols by assay HPLC chromatography of which 035.5% of hydroxycinnamic acids (chlorogenic acid) ° 26.9% of flavonols (quercetin) ° 24.3% of dihydrochalcones (phloridzin) 013.3% of flavan-3ols (catechins and catechin polymers) Example 3: Preparation of an apple extract composition A volume of 7 tons of dried apple seeds is shelled and separated to obtain 1.4 tons of shells. These shells are dry blended with 0.7 tons dried apple skin fraction isolated from a whole dry pomace. The mixture is loaded into a tank and extracted with a total volume of 47900 liters of hot water (60 ° C). The mixture is separated on a horizontal decanter and clarifier clarifier to obtain 43 700 liters of clear juice. The juice is acidified to pH 2.5 with a citric acid solution.

The acidified liquid is ultrafiltered on an installation of 50 KD organic spiral membranes to recover 41,500 liters of permeate. The permeate is passed through a column of 200 liter adsorbent resin previously equilibrated by passing 2 volumes (400 liters) of a 0.5% citric acid solution. The column, in which the polyphenols are retained, is washed with 2 V of 0.5% citric acid solution and then washed with 2V of water to remove the residual acid. The polyphenols are removed by passing 1.5V of 70% ethanol followed by a shoot with 1.5V of water. The total alcohol mixture (600 liters) is distilled to obtain 33 kg of concentrate. The concentrate is dried on a single atomization tower to recover 5.2 kg of powder. The analyzes give 35% of total polyphenols by the method of Folin and 30% of total polyphenols by assaying HPLC chromatography of which: - 15% - 21.4% of hydroxycinnamic acids (chlorogenic acid) - 35% of flavonols (quercetin) - 27, 4% dihydrochalcones (phloridzine) - 15.2% flavanols (catechins and catechin polymers)

EXAMPLE 4 Preparation of an apple composition The same mixture of raw materials and the same extraction method as in Example 3 are used. The clear juice from the clarifier (de-worming centrifuge) is glued with gelatin at 0.3 g per liter to remove some of the procyanidol polymers (tannins). The resulting suspension is centrifuged to remove most of the gelatine / tannin complex floc. The centrifuged product is acidified and ultrafiltered to remove residual colloids. The ultrafiltration permeate is passed over the 200-liter column previously equilibrated at acidic pH. The procedure is then as in Example 3 to obtain 4.8 kg of powder, the analysis of polyphenols gives 42% total polyphenols by Folin colorimetric assay and 36% total polyphenols by HPLC chromatography of which: - 27.8 % of hydroxycinnamic acids (chlorogenic acid) - 32.5% of flavonols (quercetin) - 33% of dihydrochalcones (phloridzine) - 6.7% of flavanols (catechins and catechin polymers) Note that taken separately, the 3 groups chlorogenic acid, quercetin and phloridzine are almost equivalent in amount.

EXAMPLE 5 The aim of this test is to obtain an extract which is very rich in hydroxycinnamic acids (chlorogenic acid). Highly clarified and cider apple juice is adjusted to pH 7 with dilute sodium hydroxide. The basic extract, 100 liters, is passed through a column of three liters of resin previously equilibrated to pH 7. The eluate rich in phenolic acids is collected. The column is washed with 2 volumes of neutralized water with diluted sodium hydroxide. The eluate and the wash water are collected mixed and acidified to pH 2.5 with citric acid and then passed on a second column of adsorbing resin of 3 liters previously acidified. 2 volumes of citric acid solution are passed to remove undesirable elements. The two columns are washed separately with 1.5V of water each. The polyphenols of the basic column are recovered separately with two levels of 1.5% alcoholic solutions of 40% alcohol followed by 1.5 V of water and then 1.5 V of 70% alcohol followed by 1.5 V of alcohol. water. The polyphenols in the acid column are collected at once with 1.5V of 20% alcoholic solution followed by 1.5V water. The various alcoholic extracts are distilled, concentrated, frozen and dried separately by lyophilization to obtain an extract rich in hydroxycinnamic acids, an extract rich in catechins and procyanidols and a third extract rich in phloridzin and quercetin. The analysis of the polyphenols reveals that the hydroxycinnamic extract (acid polyphenols) contains 25% of total polyphenols including 66% of chlorogenic acid by HPLC assay.

The first basic polyphenol extract (40% alcohol extract) contains 35% total polyphenols including 54% flavan-3ols (catechins and procyanidols). The second basic polyphenol extract (70% alcohol) contained 18% total polyphenols including 69% dihydrochalcones (phloridzine). A mixture of different proportions of these extracts makes it possible, in a desired and controlled manner, to obtain different preparations which are rich and balanced in chlorogenic acid, quercetin, phloridzine and catechin derivatives. Extracts rich in each of the polyphenolic groups can thus be obtained as can be obtained in which the 4 groups can be equivalently distributed 14.

Example 6: Preparation of an apple extract composition, by separation of polyphenol types by chromatoqraphy and selective elution in steps In this example, the stepwise elution consists of passing the apple juice over the adsorbent column to fix all the polyphenols, then gradually remove the different homogeneous groups by passing increasing solutions of solvents. A volume of 50 liters of unclarified cider juice is batch mixed in a 70 liter basin with 3 liters of XAD adsorbent resin. The pH of the suspension is controlled and adjusted to 2.5 with a citric acid solution. The resin / juice mixture is stirred slowly for 30 minutes. Stirring is then stopped and the suspension is allowed to stand for 25 minutes for the resin / polyphenol complex to settle to the bottom of the basin. The maximum of the supernatant is removed and acidulated water is added for a second phase of slow stirring followed by static decantation. These batch steps make it possible to eliminate the colloidal elements that can clog a chromatography column (fermentation yeasts, cell debris, proteins, polysaccharides, etc.). This batch stirring step can be avoided by clarifying the acidified juice by filtration, microfiltration or ultrafiltration. After removing the second supernatant, the slurry resin suspension is loaded into a 3 liter glass column equipped at its base with a porous sinter. The resin column is washed and stabilized with 6 liters (2V) of acidulated water and then 2V of water. The different polyphenolic groups are eluted by the alternating passage of increasing concentrations of ethanol (5%, 10%, 20%, 40%, 70%) and washing with water. The volumes of diluted alcohol and water correspond to 2 volumes of resin (6 liters). Alcoholic eluates are collected separately to identify their polyphenolic composition. Analyzes by HPLC show that the 5, 10 and 20% ethanol extracts contain mainly chlorogenic acid (about 50% of the total polyphenols) and are free of flavonols (quercetin) and dihydrochalcones (phloridzin). The 40% ethanol extract contains more than 76% catechins and procyanidins. The extract obtained with 70% of ethanol consists of more than 84%, is only dihydrochalcones and flavonols of which more than 59% dihydrochalcones and more than 25% flavonols. The extracts at 5, 10, 20 and 70% of ethanols can be mixed according to the proportions of chlorogenic acid, quercetin and phloridzine desired in the final mixture. Thus, an extract very rich in chlorogenic acid can be obtained as it is possible to form very balanced formulas of the order of 1A per substance as in Example 6.

Example 7: Evaluation of an apple extract composition on postprandial glucose In medical practice, the glucose test is the oral administration of glucose to determine how quickly it is released into the blood.

A. Protocol The glucose tolerance test is carried out during a morning (07h to 30h) since glucose tolerance has a diurnal rhythm with a significant decrease in the afternoon. Volunteers must be fasting during the previous night of 8-14 hours (water is allowed). On an empty stomach, at zero time before ingestion of glucose test, a measurement of blood sugar (blood sugar) is made.

The volunteer then ingests a glucose solution within 5 minutes. The blood glucose is then measured at regular intervals. The products tested are a control solution of 75 g of pure glucose in 300 ml of water, as well as a 300 ml test solution containing 75 g of glucose and 896 mg of total polyphenols. Based on these total polyphenols, tested apple extract was equilibrated to chlorogenic acid, quercetin and phloridzine with 17.4% hydroxycinnamic acids (chlorogenic), 20.8% dihydrochalcones (phloridzin), 17.7% flavonols (quercetin). The remaining phenolic compounds are 44.1% flavan-3-ol (catechins and their polymers). Eleven volunteer candidates participated in the test. At the end of the test and depending on the results, two volunteers were discarded for their too discordant results probably related to too much physical activity during the 3 hours of tests. The analyzed results therefore relate to 9 people.

B. Results Profile of the evolution curves of blood glucose The results show little difference between the blood glucose levels measured for each of the volunteers. During the first 90 minutes, the evolution of the blood glucose curve is similar for the two products tested (placebo and test). For the control product (glucose alone) and after 3 hours, the curve falls back to the starting fasting level. In the case of the test product containing the polyphenols, the curve does not return to the fasting blood glucose level even after 3 hours after ingestion.

Speed of use of glucose From 90 minutes after ingestion of drinks, apple extract tested seems to limit the rate of use of glucose. This results in a 33% decrease in the rate of disappearance of postprandial blood sugar.

Comparison of the areas under the evolution curves of the glycemia It is noted that for the dose of apple extract tested in relation to 75 g of glucose ingested, there is no significant effect on the glycemia during the first 90 minutes of postprandial kinetics. However, since the apple extract decreases the rate of glucose utilization beyond 90 minutes, the areas and relative amounts of glucose remain higher for the periods 90-120 minutes and 120-180 minutes. So, overall, it is found that blood glucose is higher for the periods farthest from the time of ingestion of the product.

It is known that even in the postprandial period, glucose metabolism is managed by a coordinated set of physiological responses. The two important parameters are first and foremost the regulation of the release of the food sugar into the blood pool, resulting in a maximum rate of sugar absorption (peak blood glucose) and secondly, the rate of assimilation or provision of glucose in the tissues. Thus, up to 30 minutes after ingestion of the food, the increase in blood glucose can be generally attributed to an increase in its release in the blood following the increase in intestinal absorption. Once the maximum peak is reached, the peripheral metabolism of glucose plays an increasingly important role. Deliveries in muscle and adipose tissue and liver metabolism will then have a significant effect on total plasma concentration. As a result, the rates observed above the maximum peak of blood glucose (beyond 30 minutes) can not be attributed solely to intestinal absorption alone. The results obtained here are particularly specific and different from those observed with other herbal extracts known to regulate blood glucose. Thus, studies on coffee extracts give often contradictory results. High coffee consumption or ingestion of pure caffeine would tend to increase blood glucose levels as early as the first 25 minutes of the postprandial period (Battram et al., 2006, J. Nutr., 136: 1276-1280; et al., 2004, J. Nutr.134: 2528-2533, Lane et al., 2004, Diabetes Care, Vol 27, No. 8, 2047-2048). On the other hand, decaffeinated coffee would give a lowering of the blood glucose curve and lower total blood sugar volumes (Battram et al., 2006, J. Nutr 136: 1276-1280). The results of the present study are closer to those obtained in some studies with tea extracts. Overall, the consumption of the extract does not give significant differences from the control during the first hour after ingestion of the product. On the other hand, beyond the hour (in the zone of 60 to 70 minutes) one can observe a decrease in the blood glucose curve below the control followed by a rise above 120 minutes (Bryans et al., 2007, Journal of the American College of Nutrition, Vol 26, No. 5, 471-477). This study shows that apple extract particularly regulates the last phase of postprandial glucose, ie when peripheral metabolism intervenes (hepatic metabolism, muscular uses, lipid accumulation in fatty tissues). Thus, the favorable effect of the extract can not be explained by the only inhibition of the intestinal absorption of sugar. This regulation probably manifests itself in the secretion of insulin by pancreatic B-cells and the action of digestive-type hormones such as GIP (glucose-io-dependent insulinotropic-polypeptide) and GLP-1 (glucagon-like peptide). -1). This late effect of the extract could be related to the time required for the extract's active ingredients or their metabolites to be absorbed. It is known, for example, that certain phenolic compounds such as catechins appear in the blood 30 minutes after their ingestion and the maximum peak is reached at about 60 minutes. It is interesting to note that maintaining a higher level of circulating glucose during the last phase of glucose could be beneficial in the context of intense and prolonged efforts requiring a prolonged supply of energy.

Example 8: Comparative Trial of the Activities of an Apple Extract Composition of the Invention with a Phloridzine-enriched Apple Extract Composition The object of this test was to show that phloridzine is not the dominant component providing the desired effects on blood sugar, lipogenesis and the risk of gaining weight. The aim was to evaluate the effects of two extracts of different compositions, particularly in terms of phloridzin levels, on the differentiation of stem cells into adipocytes. It is known that one of the undesirable consequences of the excessive assimilation of sugars by the cells is its storage through the synthesis and accumulation of lipids. The resultant logic of this lipid accumulation is the formation of important layers of adipose tissue, weight gain and obesity. The expected result in this test is therefore to verify that the apple extracts are capable of blocking the synthesis and accumulation of lipids and that the importance of this inhibition is related to the composition of the extract. More specifically, the test aims to show that the extracts richest in phloridzine do not necessarily give the best results. The parameters studied are cell morphology and lipid accumulation.

A. Materials and Methods Morphological monitoring of differentiation and intracellular lipid accumulation was performed.

Cells Used The cells used in culture are human mesenchymal poietic stem cells (hMSC). A cytotoxicity test is carried out beforehand on undifferentiated cells in order to select the tolerated concentrations and therefore to be tested. Cell culture conditions The cells were cultured in a cell culture incubator in an atmosphere of 5% CO2 at 37 ° C and 95% relative humidity. Cells were inoculated into 3 96-well cell culture plates at 20,000 hMSC cells per well in growth medium. [Reference Medium, SVP] After incubation for 3 days at 37 ° C, the culture medium was removed and replaced with a hMSC cell differentiating medium to adipocytes [Reference Medium, SVP]. This differentiation medium contained or not the apple extracts tested. After the culture time, the morphological differentiation into adipocytes is followed on the cell mats of the plates using a fluorescent marker specific for intracellular lipids. For easy observation, representative photographs were made using a camera of the NIKON DX-1200 type.

Qualitative and quantitative constitution of the tested composition Extract 1: rich and balanced in all four groups of phenolic compounds of apple This powder extract contained 41% of total polyphenols assayed by colorimetry according to the Folin method. Assayed by HPLC chromatography, the following distribution is obtained: 15% of flavanols (catechins and their procyanidol polymers), ie 36.6% of the total polyphenols 8.5% of flavonols (quercetin), ie 23.4% of the polyphenols total of 8.7% of hydroxycinnamic acids (chlorogenic acid), ie 21.2% of the total polyphenols 5% of dihydrochalcones (phloridzin), ie 12.2% of the total polyphenols

Extract 2: Highly enriched in phloridzine This powder extract contained 42% of total polyphenols by colorimetry and 35% of dihydrochalcones (phloridzine) by HPLC chromatography. This indicates that phloridzin accounts for 83.3% of the total polyphenols in this extract. It should be noted that this extract is almost free of chlorogenic acid, quercetin and catechin derivatives.

B. Results Pre-cytotoxicity The cytotoxicity results and the morphological observation of the cells as a function of the concentrations of the two extracts showed that the cultures were more stable in the presence of the balanced extract 1 in different phenolic compounds, compared with the extract 2 strongly enriched in phloridzine. The high concentration of phloridzine therefore seems to bring greater toxicity. This may be related to the property of depolarization of cell membranes by phloridzin. These observations led to the selection of the concentrations used for the rest of the test, namely 0.04 mg / ml of the balanced extract 1 and 0.02 mg / ml of the phloridazine-rich extract 2. Note that despite these two different concentrations, extract 2 tested at 0.02 mg / ml provides much more phloridzine (7 mg / ml) compared to the amount (2 mg / ml) provided by 0.04 mg / ml From extract 1. Overall extract 2 brings 3.5 times more phloridzin.

Adipogenesis Compared with the differentiated control, the 11-day cell cultures show that the two extracts tested are capable of regulating the morphological modification of the cells and especially of inhibiting the differentiation into adipocytes as well as the accumulation of intracellular lipids. However, the extract 1 rich in chlorogenic acid and balanced in different phenolic compounds is the one that gives the best result. These results show that the inhibition of cell differentiation, lipogenesis and lipid accumulation is not correlated with a high concentration of the phloridzin extract. The other phenolic compounds, especially chlorogenic acid and quercetin, seem to play a predominant role. These results thus clearly confirm that phloridzine is not the essential active component and is not the only element to be taken into account in explaining the effects of the apple or its extracts, especially on the cellular assimilation of sugars, synthesis and lipid accumulation. A synergistic action of the different compounds seems more appropriate.

Example 9: Effect of an apple extract composition on the activity of the different isoforms of nitric oxide synthetases Nitric oxide (NO) is one of the cellular mediators of physiological and pathological processes. NO is synthesized from arginine by the enzymes synthetases of nitric oxide (NOS). There are three isoforms called endothelial (eNOS), neuronal or central (nNOS or cNOS) and inducible (iNOS). The first two are constitutive in the body, while the third (iNOS) is an inducible enzyme strongly expressed by the inflammatory process. While small amounts of NO from the constituent isoforms (eNOS and cNOS) are essential for the proper functioning of the body, a significant increase in the NO synthesized by iNOS participates in triggering and amplifying the inflammatory process. Compounds capable of regulating the synthesis of NO by the three isoforms and more particularly of inhibiting the synthesis of NO by iNOS, are more and more studied for their anti-inflammatory effects. Similarly, substances promoting moderate NO production by eNOS and nNOS, more particularly eNOS, are potential means of protecting the body. Endothelial nitric oxide (eNO) and the enzyme that synthesizes this molecule (eNOS) are important for vasodilatation, blood pressure reduction, better oxygen circulation, reductions in hypertension and risks. cardio-vascular diseases. The purpose of this test is therefore to evaluate the effects of different concentrations of the tested extract on the expression of the genes of the various NO synthetases, ie the so-called central nitric oxide synthetase I (cNOS). , nitric oxide synthetase II called inducible (iNOS) and so-called endothelial nitric oxide synthetase III (eNOS).

A. Protocol The tested product is extract 1 of Example 8. The test is carried out on aged and senescent normal human dermal fibroblast cultures. The simplified DNA arrays method with anti-aging theme was used. A preliminary cytotoxicity test was performed to determine tolerable extract doses that could be used for the gene expression efficiency test. The analyzes are carried out on cell cultures of 6 hours and 24 hours. The results are expressed in relative units of expression (ER) relative to a control gene.

B. Results The preliminary cytotoxicity test made it possible to select the following concentrations for the continuation of the study: 0.01 mg / ml; 0.005 mg / ml and 0.0005 mg / ml The results, in terms of gene expression level of the 3 isoforms, are summarized in Table 1 below with respect to the control (100).

Table 1 ISOFORM CONCENTRATION OF APPLE EXTRACT

0.01 mg / ml 0.005 mg / ml 0.0005 mg / ml 6H 24H 6H 24H 6H 24H

cNOS 77 63 98 49 72 69 1NOS 100 100 100 100 100 100 35 eNOS 164 133 147 107 106 111 The results show that, whatever the cell culture time (6 or 24 hours) and the concentration of apple extract, the expression of inducible NO synthase (iNOS) is rigorously stable. It is interesting to note that the expression of the gene of this enzyme involved in the inflammatory process is not at all influenced by the presence of the apple extract in the culture medium. This would indicate that the extract is potentially non-inflammatory. It is found that the central NO synthase (cNOS) gene is repressed by the presence of apple extract. But this inhibition is very moderate and is not correlated with the concentration of the apple extract. Inhibition is more accentuated by the duration of cell cultures. At 6 hours of culture there is only 17.7% average rate of inhibition against 40% at 24 hours. The endothelial NO synthase (eNOS) gene is the most sensitive to the presence and concentration of apple extract. The observed results also depend on the duration of the cell cultures. At 6 hours of cell culture, it is found that the gene is overexpressed and that this increase is correlated with the concentration of the apple extract. Thus, respectively 6%, 47% and 64% increase for 0.0005 mg / ml, 0.005 mg / ml and 0.01 mg / ml of apple extract. At 24 hours of cell culture, only a weak overexpression of the eNOS gene is observed. In addition, this slight overexpression is no longer correlated with the concentration of apple extract, and only the highest concentration of apple extract gives a noticeable result. It is interesting to note that of the three NOS isoforms evaluated, only the eNOS involved in the synthesis of endothelial nitric oxide used in vasodilation and the reduction of blood pressure and therefore of hypertension is overexpressed. In addition, this overexpression of the eNOS gene remains relatively moderate. This is in line with other authors' observations stressing that a moderate synthesis of eNO is required, since excessive NO production would, on the contrary, lead to toxic effects.

Example 10 Composition for topical application The composition for topical application consists of a dermatological gel slimming anti-cellulite. The extract used in this example is that obtained in Example 2.

The qualitative and quantitative constitution of the composition is described below.

INGREDIENTS PROPORTIONS (%) PHASE 1 Demineralized water 65,33 Carbopol Ultrez 21,00,50 Sodium hydroxide (20% solution) 00,04 PHASE 2 Ethoxydiglycol oleate 16,00 Phenonip 00,60 Vitamin E acetate 00,30 PHASE 3 Demineralized water 10 , 00 Extract from example 2 05.00 PHASE 4 Novemer EC-1 01.00 Apple seed oil 00.50 PHASE 5 Yellow 6 00.20 Fragrance 00.40 Sodium hydroxide (20%) 00.13 Process of preparation of the composition - prepare the NaOH solution in demineralised water - prepare phases 2, 3 and 4 - sprinkle Carbopol in water (65.33 g), when well hydrated, the mixture at low speed - neutralize with NaOH - add phase 2 to phase 1 - add phase 3 with slow stirring until uniform - add phase 4 stirring until uniform - add ingredients from phase 5 order by shaking until uniform

Example 11: Composition for oral administration in the form of a dietary supplement The composition for oral administration consists of anti-cellulite slimming capsules.

The extract used is that of Example 2 The qualitative and quantitative constitution of the composition is described below. INGREDIENTS PROPORTIONS

Extract of Example 2 135.00 mg Vitamin D (cholecalciferol) 70.00 IU 20 Magnesium stearate 0.5 mg Pomelite (apple fiber) QSP 400 mg

Example 12: Composition for oral administration in the form of a beverage The composition for oral administration consists of an aromatized mineral water with a slimming effect. The extract used is that in the acidic polyphenol fraction of Example 4 The qualitative and quantitative constitution of the composition is described below. 30 INGREDIENTS PROPORTIONS Acidic extract of Example 4 00.40 Carotene (powder 10%) 00.15 Vitamin C (powder) 00.20 35 Citric acid 00.25 Vitamin E (powder 50%) Concentrated apple juice (70 Brix ) Anthocyanin P color Apple aroma (powder) Lemon aroma (powder) Water QSP00,10 07,00 00,01 00,02 00,01 100 g Example 13: Composition for oral administration in the form of sports drink The composition for administration oral consists of a drink for energy management and protection against oxidative stress caused by muscular effort, especially for athletes. This composition can be ingested before, during or after the muscular effort. The qualitative and quantitative constitution of the composition is described below.

INGREDIENTS PROPORTIONS

Apple juice 20 Concentrated juice Maltodextrins Pomeline (apple oligosaccharides) Pomelite (apple polysaccharides) Pectin HM 25 Pomactiv Shape (apple polyphenols) Pineapple aroma Vitamin C Vitamin E Caffeine 30 Niacin Vitamin B-6 Vitamin B-2 Vitamin B-1 Vitamin B-12 35 237.624 g 027,000 g 012,000 g 009,000 g 009,000 g 003,000 g 001,500 g 000,600 g 1,000,150 g 000,050 g 000,045 g 000,025 g 000,0025 g 000,002 g 000,002 g 000,008 mg TOTAL 300,000 30

Claims (11)

REVENDICATIONS1. A pome fruit extract composition of the Rosaceae family comprising at least 25 percent by weight of polyphenols based on the total dry weight of said composition, said composition having a content of at least 15 percent by weight of chlorogenic acid and said composition comprising a combination of: - at least 15 percent by weight of flavonols (quercetin), - at least 10 percent by weight of dihydrochalcones (phloridzin), and Io - at least 10 percent by weight of flavanols (catechins), the weight percentages of chlorogenic acid, flavonols, dihydrochalcones and flavanols being expressed relative to the total weight of the polyphenols. 15 2. Composition according to claim 1, characterized in that it comprises at least 20, 25, 30, 35, 40, 45, 50 or 55 percent by weight of chlorogenic acid, relative to the total weight of the polyphenols contained in said composition 3. Composition according to any one of claims 1 and 2, characterized in that it comprises at least 15, 20, 25 or 30 percent by weight of flavonols, including quercetin, relative to the total weight of the polyphenols . 4. Composition according to any one of claims 1 to 3, characterized in that it comprises at least 15 or 20 percent by weight of dihydrochalcones, including phloridzine, relative to the total weight of polyphenols. 5. Composition according to any one of claims 1 to 4, characterized in that it comprises at least 15 or 20 percent by weight of flavanols, including catechins, relative to the total weight of polyphenols. 6. Composition according to any one of claims 1 to 5, characterized in that it comprises at least 30, 35, 40, 45, 50, 55, 60, 65, 70 or 75 percent by weight of total polyphenols, relative to the total weight of dry matter of said composition. 35 7. A composition according to claim 1, characterized in that it comprises at least 25 percent by weight of total polyphenols, for example from 30 to 80 percent by weight of total polyphenols, relative to the total weight of dry matter of the composition, and comprises: - at least 15 percent by weight, for example from 30 to 60 percent by weight, of hydroxycinnamic acids (chlorogenic acid), - at least 15 percent by weight, for example from 25 to 35 percent by weight, one hundred percent by weight of flavonols (quercetin), at least 10 percent by weight, for example from 20 to 30 percent by weight, of dihydrochalcones (phloridzin and other phloretin derivatives), and at least 10 percent by weight by weight, for example from 15 to 25 percent by weight, of flavanols (catechins), the percentages by weight of chlorogenic acid, flavonols, dihydrochalcones and flavanols being expressed relative to the total weight of the polyphenols. 8. A composition according to claim 1, wherein the polyphenols of interest are present in the following contents: 20 to 30 percent by weight of chlorogenic acid, 20 to 30 percent by weight of flavonols, and quercetin, 20 to 30 percent by weight of dihydrochalcones, including phloridzine, and 20 to 30 percent by weight of flavonols, including catechins, the percentages by weight being based on the total weight of said polyphenols of interest in the composition. 9. Food, dietary, cosmetic, nutraceutical or cosmeceutical composition comprising a pome fruit extract composition of the family Rosaceae according to one of claims 1 to 8, in combination with one or more physiologically compatible vehicles. 10. A process for the manufacture of a pome fruit extract composition of the Rosaceae family, comprising the following steps: a) contacting the starting raw material in solution in a suitable solvent on an adsorbent resin of the styrene-divinylbenzene type, b) eluting a first fraction enriched in hydroxycinnamic acids with an aqueous solvent comprising from 15% to 25% by weight of ethanol, and recovering said first fraction, and then obtaining a powder of said first fraction by removal of the aqueous solvent, c) elution of a second fraction enriched in flavanols with an aqueous solvent comprising from 35% to 45% by weight of ethanol, and recovery of said second fraction, and then obtaining a powder of said second fraction by removal of the aqueous solvent, d) eluting a third fraction enriched in dihydrochalcones and flavonols with an aqueous solvent comprising from 60% to 80% by weight of ethan ol, and recovering said third fraction, then obtaining a powder of said third fraction by removing the aqueous solvent, e) preparing a composition of pome fruit extract of the Rosaceae family according to any one of Claims 1 to 8 by mixing the powders obtained in steps b), c) and d) respectively, in appropriate proportions. 11. A process for the manufacture of a pome fruit extract composition of the Rosaceae family comprising the following steps: a) preparing a fraction enriched in hydroxycinnamic acids according to a process comprising the following steps: a1) placing contacting the starting raw material in solution in a suitable solvent and adjusted to an acidic pH on a styrene-divinylbenzene adsorbent resin also adjusted to an acid pH, the acidic pH being from pH 2 to 3.0, a2) eluting a first fraction enriched in hydroxycinnamic acids with an aqueous solvent comprising from 30% to 50% by weight of ethanol, and recovering said first fraction, then obtaining a powder of said first fraction by removing the aqueous solvent, b) preparing a second fraction enriched in flavanols and a third fraction enriched in flavonols and dihydrochalcones, according to a process comprising the following steps: b1) contacting the starting raw material in solution in an appropriate solvent and adjusted to a basic pH on an adsorbent resin of the styrene-divinylbenzene type also adjusted to basic pH, the basic pH consisting of a pH ranging from 6.5 to 7.5, b2) eluting a second fraction enriched with flavanols, with an aqueous solvent comprising from 35% to 45% by weight ethanol, and recovering said second fraction, and then obtaining a powder of said second fraction by removal of the aqueous solvent, b3) elution of a third fraction enriched in flavonols and dihydrochalcones, with an aqueous solvent comprising from 65% to 75% by weight of ethanol, and recovery of said third fraction, then obtaining of a powder of said third fraction by removal of the aqueous solvent; c) preparation of a pome fruit extract composition of the Rosaceae family according to any one of claims 1 to 8, by mixing the powders obtained respectively in steps a) and b), in appropriate proportions. 15