Use of polyphenols and polyphenol-based compositions to improve the oral bioavailability of hydroxytyrosol - Patent Application 20070233334

Abstract

The present invention relates to a composition of natural origin, comprising hydroxytyrosol and a polyphenol other than hydroxytyrosol that does not naturally occur in the olive tree (O. europaea), which, once orally administered, allows for particularly high bioavailability of hydroxytyrosol and particularly high anti-atherosclerotic activity of hydroxytyrosol. The composition may comprise a mixture of plant extracts. The present invention also relates to the use of the composition for the treatment of cardiovascular diseases, particularly atherosclerosis.

Description

[Technical field]

[0001] The present invention relates to a polyphenol composition comprising hydroxytyrosol for increasing the bioavailability of hydroxytyrosol and increasing the therapeutic activity of hydroxytyrosol in the prevention and treatment of cardiovascular diseases, more specifically atherosclerosis, cerebrovascular accidents, peripheral vascular disease, and coronary disease. [Background technology]

[0002] Cardiovascular disease (CVD) remains the leading cause of death worldwide, with approximately 17.8 million deaths due to CVD in 2017. This represents a 12.5% ​​increase in mortality over the past decade.

[0003] Atherosclerotic vascular disease (ASVD), or atherosclerosis, is one of the most important causes of CVD. It is a chronic vascular inflammatory disease associated with oxidative stress and endothelial dysfunction, in which oxidized low-density lipoprotein (oxLDL) has been suggested to play an important role. Because oxLDL binds to multiple receptors with higher affinity than native LDL, it is absorbed by macrophages more quickly, which leads to cholesterol accumulation and atherosclerotic plaque formation. Furthermore, oxLDL promotes endothelial cell activation and dysfunction, vascular smooth muscle cell migration and proliferation, and platelet activation, causing inflammatory responses, endothelial dysfunction, and exacerbating the atherogenic process. As a result, the concentration of circulating oxLDL is considered one of the most important biomarkers of atherosclerosis progression.

[0004] In general, the treatment of atherosclerosis consists of changing lifestyle habits to healthier ones. However, when this is insufficient, it is usually combined with the administration of drugs, which are most often aimed at lowering blood pressure and / or cholesterol. In advanced cases, surgical procedures may be required to widen the narrowed artery / arteries. Therefore, the current state of the art requires a procedure that allows the benefits of a healthy lifestyle to outweigh the development of atherosclerosis. Summary of the Invention

[0005] The present invention is based on the finding that by combining hydroxytyrosol (HT) present in olive oil with polyphenols not naturally present in the olive tree (Olea europaea) or in olive oil, such as those present in almond skin extract, grapefruit, flax or knotweed extract, it is possible to increase the bioavailability of HT after oral administration of HT compared to when HT is administered without polyphenols. Furthermore, the inventors observed how administration of a composition comprising HT and almond skin polyphenols (ASP) to subjects with moderate hypercholesterolemia (treatment group) reduces serum levels of oxidized low-density lipoprotein (oxLDL) and the oxLDL / LDL ratio. However, in subjects with moderate hypercholesterolemia treated with placebo (control group), serum levels of oxLDL are elevated after treatment, as is the oxLDL / LDL ratio.

[0006] Thus, a first aspect of the present invention relates to the use of at least one polyphenol which is not hydroxytyrosol and / or does not naturally occur in the Olea europaea (O. europaea) tree, for increasing the bioavailability of hydroxytyrosol following oral administration of hydroxytyrosol or for increasing the anti-atherosclerotic effect of hydroxytyrosol following oral administration of hydroxytyrosol.

[0007] In a second aspect, the present invention relates to a polyphenol which is not hydroxytyrosol and / or does not naturally occur in the O. europaea tree, for use in increasing the bioavailability of hydroxytyrosol after oral administration or in increasing the anti-atherosclerotic effect of hydroxytyrosol after oral administration.

[0008] In a third aspect, the present invention relates to hydroxytyrosol for use in the treatment of atherosclerosis, administered orally in combination with a polyphenol which is not hydroxytyrosol and / or does not naturally occur in the O. europaea tree.

[0009] In a fourth aspect, the present invention relates to a composition or kit of parts comprising hydroxytyrosol and at least one polyphenol, wherein the polyphenol is not hydroxytyrosol and / or the at least one polyphenol does not naturally occur in the O. europaea tree.

[0010] In a fifth aspect, the present invention relates to a method of obtaining a composition according to the fourth aspect of the invention, comprising contacting a composition comprising hydroxytyrosol with a composition comprising at least one polyphenol which is not hydroxytyrosol and / or does not naturally occur in the O. europaea tree.

[0011] In a sixth aspect, the present invention relates to a pharmaceutical composition comprising the composition of the fourth aspect of the invention or the composition obtainable by the method of the fifth aspect of the invention, and a pharma- ceutically acceptable excipient.

[0012] In a seventh aspect, the present invention relates to a food supplement or dietary supplement comprising the composition of the fourth aspect of the invention or the composition obtainable by the method of the fifth aspect of the invention and a nutritionally acceptable excipient.

[0013] In an eighth aspect, the present invention relates to a composition or a kit of parts according to the fourth aspect of the invention, a composition obtainable by the method according to the fifth aspect of the invention, a pharmaceutical composition according to the sixth aspect of the invention, or a food supplement or dietary supplement according to the seventh aspect of the invention, for use in medicine.

[0014] In a ninth aspect, the present invention relates to a composition or a kit of parts according to the fourth aspect of the invention, a composition obtainable by the method according to the fifth aspect of the invention, a pharmaceutical composition according to the sixth aspect of the invention, or a food supplement or dietary supplement according to the seventh aspect of the invention, for use in the treatment of cardiovascular disease. [Brief description of the drawings]

[0015] [Figure 1] FIG. 1 is a diagram summarizing the study. [Diagram 2] Figure 1 shows the oxLDL-cholesterol / LDL-cholesterol ratio at the beginning, 4 weeks, and 8 weeks after treatment in subjects who received placebo or extract. Mean values ​​are represented by dots (dark grey for control group, light grey for extract group) and standard errors are represented by vertical lines. Different letters indicate significant differences between each time point for each group. Asterisks (*) indicate p<0.05 between groups at the corresponding time point. [Diagram 3] Figure 3A is a chromatogram at 280 nm separating olive wood extract polyphenols (bottom chromatogram) and almond extract polyphenols (top chromatogram) and Figure 3B is a chromatogram at 330 nm separating olive wood extract polyphenols (top chromatogram) and almond extract polyphenols (bottom chromatogram). [Figure 4] Figure 4A is a chromatogram at 280 nm separating olive wood extract polyphenols (top chromatogram) and grapefruit extract polyphenols (bottom chromatogram) and Figure 4B is a chromatogram at 330 nm separating olive wood extract polyphenols (top chromatogram) and grapefruit extract polyphenols (bottom chromatogram). [Diagram 5] Figure 5A is a chromatogram at 280 nm separating olive wood extract polyphenols (top chromatogram) and flax extract polyphenols (bottom chromatogram) and Figure 5B is a chromatogram at 330 nm separating olive wood extract polyphenols (top chromatogram) and flax extract polyphenols (bottom chromatogram). [Figure 6] Figure 6A is a chromatogram at 280 nm separating olive tree extract polyphenols (top chromatogram) and Polygonum decipiens extract polyphenols (bottom chromatogram). Figure 6B is a chromatogram at 330 nm separating olive tree extract polyphenols (top chromatogram) and Polygonum decipiens extract polyphenols (bottom chromatogram). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0016] The inventors have observed that several groups of polyphenols, other than HT, that are not naturally found in the O. europaea tree, increase the bioavailability of HT following oral administration of HT. Furthermore, administration of a composition containing HT and almond skin polyphenols to subjects with moderate hypercholesterolemia reduces serum levels of oxLDL.

[0017] Use of polyphenols other than hydroxytyrosol for increasing the bioavailability and anti-atherosclerotic activity of hydroxytyrosol, and use of hydroxytyrosol in the treatment of atherosclerosis by administering hydroxytyrosol together with non-hydroxytyrosol polyphenols

[0018] In a first aspect, the present invention relates to the use of at least one polyphenol that is not hydroxytyrosol and / or that does not naturally occur in the Olea europaea (O. europaea) tree for increasing the bioavailability of hydroxytyrosol after oral administration of hydroxytyrosol or for increasing the anti-atherosclerotic effect of hydroxytyrosol after oral administration of hydroxytyrosol. Alternatively, the first aspect of the present invention relates to a method for increasing the bioavailability of hydroxytyrosol after oral administration of hydroxytyrosol or for increasing the anti-atherosclerotic effect of hydroxytyrosol after oral administration of hydroxytyrosol, the method comprising administering at least one polyphenol that is not hydroxytyrosol and / or that does not naturally occur in the Olea europaea (O. europaea) tree.

[0019] In a second aspect, the present invention relates to a polyphenol which is not hydroxytyrosol and / or does not naturally occur in the O. europaea tree, for use in increasing the bioavailability of hydroxytyrosol following oral administration of hydroxytyrosol or for increasing the anti-atherosclerotic effect of hydroxytyrosol following oral administration of hydroxytyrosol.

[0020] In a third aspect, the present invention relates to hydroxytyrosol for use in the treatment of atherosclerosis, wherein the hydroxytyrosol is orally administered in combination with a polyphenol that is not hydroxytyrosol and / or does not naturally occur in the O. europaea tree. Alternatively, the third aspect of the present invention is defined as a method of treating atherosclerosis in a patient, the method comprising orally administering to the patient hydroxytyrosol, the hydroxytyrosol orally administered in combination with a polyphenol that is not hydroxytyrosol and / or does not naturally occur in the O. europaea tree.

[0021] As used herein, the term "hydroxytyrosol" refers to a polyphenolic compound commonly known by that name to those skilled in the art. In nature, hydroxytyrosol is present in olive trees (e.g., Olea europaea species) and is concentrated mainly in the leaves and fruit. In a particular embodiment, the term refers to the compound with the IUPAC name 4-(2-hydroxyethyl)benzene-1,2-diol.

[0022] As used in the present invention, the phrase "polyphenols that are not hydroxytyrosol and / or that do not naturally occur in Olea europaea trees" or "polyphenols that are not hydroxytyrosol and / or at least one of which is not naturally occurring in O. europaea trees" refers to polyphenols other than hydroxytyrosol as defined above (i.e. the characteristics of this polyphenol do not correspond to those of the product described in the above definition of hydroxytyrosol). Alternatively or additionally, this phrase refers to polyphenols that are not naturally synthesized by trees of the O. europaea species (i.e. without the need for human intervention in the creation and selection of trees belonging to the species or in their cultivation), regardless of the cultivation method used, preferably regardless of the variety of the O. europaea species and more preferably regardless of the individual of the species from which the extract is obtained. Thus, in a particular embodiment, polyphenols that do not naturally occur in O. europaea trees are those that are not synthesized by trees of the O. europaea species, as just described. In a particular embodiment, the tree of the O. europaea species is a wild variety of the common olive tree, i.e., the Olea europaea sylvestris variety. In another particular embodiment, the tree of the O. europaea species is of a subspecies selected from the group consisting of O. europaea europaea, O. europaea cerasiformis, O. europaea cuspidata, O. europaea guanchica, O. europaea laperrinei, and O. europaea maroccana. In another particular embodiment, the tree of the O. europaea species is a variety of any of the subspecies. In another particular embodiment, the tree is any organism included in one of the taxonomic ranks of O. europaea mentioned. In a particular embodiment, in any aspect of the invention, the species of O. europaea mentioned refers to the species O. europaea L.

[0023] In certain embodiments, polyphenols that are not hydroxytyrosol and optionally do not occur naturally in any of the Olea europaea trees are referred to in any aspect of the invention by the phrase "non-hydroxytyrosol polyphenols," "polyphenols other than hydroxytyrosol," or "polyphenols different from hydroxytyrosol."

[0024] As defined above, the method for determining whether a polyphenol occurs naturally in a tree of the O. europaea species, i.e. whether it is naturally synthesized by the tree, comprises obtaining a plant extract from at least one plant product of a tree as described in the definition and determining whether the polyphenol is present in the extract. The absence of the polyphenol in the extract indicates that the polyphenol is not found in the tree and is not synthesized by the tree. A method for obtaining a plant extract from a plant product is described below. A method for identifying the polyphenols present in a plant extract is described below. Definitions of plant extract and plant product are also provided below.

[0025] In certain embodiments, the polyphenol that is not hydroxytyrosol and / or does not naturally occur in the O. europaea tree does not naturally occur in the fruit or leaves of the O. europaea tree, and preferably the polyphenol does not naturally occur in the leaves of the O. europaea tree. As will be appreciated by those skilled in the art, methods for determining whether a polyphenol naturally occurs in the fruit and / or leaves of the O. europaea tree include the methods described above, where the plant extract is obtained from the relevant plant product, i.e., the fruit and / or leaves of the O. europaea tree.

[0026] As used in the present invention, the expression "polyphenol" refers to a group of chemicals generally known by that name to those skilled in the art. It specifically refers to chemicals found in plants, characterized by the presence of one or more phenolic rings and conjugated double bonds. Polyphenols are found mainly in conjugated form, associated with carbohydrates by binding through -OH groups, although direct binding between carbohydrates and phenolic groups also exists. It is common for polyphenols to also associate with other compounds, such as carboxylic and organic acids, amines, lipids, and even other phenolic groups. Polyphenols suitable for use in the present invention include, but are not limited to, phenolic acids (hydroxybenzoic or hydroxycinnamic acid derivatives), stilbenes, lignans, phenolic alcohols, and flavonoids, with flavonoids constituting the largest subclass. These groups of polyphenols are also used in the present invention to refer to groups of polyphenols generally known by the corresponding names to those skilled in the art.

[0027] Specifically, as used in the present invention, the term "flavonoid" refers to the largest subclass of polyphenols in the plant kingdom. The chemical structure of flavonoids consists of a diphenylpropane skeleton (C6-C3-C6) that is formed by two aromatic rings linked through three carbon atoms to form an oxygenated heterocycle (C). There are subgroups of flavonoids, and the classification of these compounds is based on the oxidation state of the heterocycle (C ring) and the position of the B ring. Within each family, there is a wide range of compounds that differ from each other in the number and position of OH groups, as well as the functional groups that may be present (methyl, sugars, organic acids). Flavonoids suitable for use according to the present invention include flavonols, flavones, flavanones, isoflavones, anthocyanidins, and flavanols (catechins and proanthocyanidins). The most representative flavonols in food are the flavan-3-ols, which can occur as anthocyanidin monomers (catechin, epicatechin, gallocatechin, and epigallocatechin), as dimers condensed with each other, or as oligomers (procyanidins), or the flavan-3-ols can occur as polymers (proanthocyanidins or condensed tannins). The compounds and classes of compounds are commonly known to those skilled in the art by the appropriate names. Non-limiting examples of flavonoids include: procyanidin, propelargonidin, prodelphinidin, catechin, epicatechin, quercetin, kaempferol, isorhamnetin, rutin, and genistein.

[0028] As used herein, the term "lignan" refers to dimeric phenolic compounds, phenylalanine derivatives, and cinnamic alcohol, whose basic structure consists of two C6C3 (n-propylbenzene) units linked by a β-β' bond.

[0029] There are several subgroups of lignans suitable for use in the present invention, such as simple lignans, complex lignans, or cyclolignans, flavolignans, coumarinolignans, stilbene lignans, or xanthone lignans, which are generally known by the relevant names to those skilled in the art. Non-limiting examples of lignans include: secoisolariciresinol diglucoside, secoisolariciresinol, matairesinoside, lariciresinol, pinoresinol, sesamin, schizandrin, deoxyschizandrin, gomisin, pregomisin, sesamolin, sesamol, sesaminol, sesamolinol, or pinoresinol.

[0030] As used herein, the term “stilbene” refers to a compound of formula C 14 H 12 The term stilbene refers to the polyphenolic compound of the formula IUPAC name trans-1,2-diphenylethylene, which exists in two isomers: trans-1,2-diphenylethylene (E-stilbene) and cis-1,2-diphenylethylene (Z-stilbene). The term stilbene includes hydroxy and alkoxy derivatives of simple stilbenes, as well as their heteroside (glycoside) forms and polymers. In certain embodiments, the term refers to the compound with the IUPAC name trans-1,2-diphenylethylene. Non-limiting examples of stilbenes include resveratrol, piceatannol, pinosylvin, pterostilbene, rhapontigenin, pterostilbene, isorhapontigenin, rhapontin, ponticin, piceid, or astringin.

[0031] As used herein, the term "phenolic acid" or "phenolic carboxylic acid" refers to a phenolic compound having a phenol ring and an organic carboxylic acid functional group (C6-C1 backbone). Non-limiting examples of phenolic acids include 3,4-dihydroxybenzoic acid, parahydroxybenzoic acid, vanillic acid, caffeic acid, paracoumaric acid, ferulic acid, syringic acid, and sinapic acid.

[0032] As used herein, the term "phenolic alcohol" refers to a phenolic compound having an alcohol functional group and a phenol functional group. Non-limiting examples of phenolic alcohols include coniferyl alcohol, sinapyl alcohol, or p-coumaryl alcohol.

[0033] In a preferred embodiment, the hydroxytyrosol used according to the invention or the polyphenols used in the invention are found as part of a plant extract.

[0034] The expression "plant extract" or "extract" as used in the present invention refers to a term commonly known to those skilled in the art. It refers to a composition containing compounds, ingredients, or substances, which are extracted from a plant organism, tree, or plant product or tissue by a method commonly known to those skilled in the art. Examples of such methods, but not limited to, include squeezing, which involves introducing the plant product into a hydraulic press to squeeze it, distillation, or cutting the plant product of interest and then extracting a sample of the contents or composition of said plant product. Another common method of obtaining a plant extract consists of treating the product or tissue from which the plant extract is to be obtained with a solvent. Examples of solvents, but not limited to, include water, ethanol, hydroalcohol, ethyl acetate, CO2, methanol, acetone, acetic acid, or hexane.

[0035] In a particular embodiment, the plant extract is obtained from fresh or dried plant products or tissues of interest using a solvent at a temperature of about 40° C. to 100° C. for 1 hour to 10 hours. In a particular embodiment, the solvent is aqueous, preferably water. In another particular embodiment, the solvent is a hydroalcohol, preferably a hydroalcohol with an alcohol content of 10% to 96%, the alcohol being preferably ethanol or methanol. The process can also include multiple extraction steps in which the plant products are extracted using the same or different solvents with the same or different alcohol content, and the obtained extracts are combined and continued in the process. In another particular embodiment, the process also includes multiple concentration steps by chromatography using a polymeric adsorption resin. The obtained extract is then treated with activated carbon, which is then removed. Finally, or alternatively, a drying step is performed to dry the purified extract.

[0036] Thus, in a particular embodiment, the method for obtaining a plant extract comprises: (i) extracting the plant product or tissue of interest from fresh or dried with a solvent at a temperature of about 40° C. to 100° C. for 1 hour to 10 hours; (ii) treating the extract obtained in step (i) with activated carbon and then removing the activated carbon; Includes.

[0037] In another particular embodiment, the method for obtaining a plant extract comprises the steps of: (i) extracting the plant product or tissue of interest from fresh or dried with a solvent at a temperature of about 40° C. to 100° C. for 1 hour to 10 hours; (ii) concentrating the resulting mixture by chromatography using a polymeric adsorption resin; (iii) treating the extract obtained in step (i) with activated carbon and then removing the activated carbon; Includes.

[0038] In a particular embodiment, step (i) of any of the above methods comprises multiple extraction steps, using the initial solvent or different solvents to extract from the plant product, and combining the resulting extracts to continue the process. If the solvent is hydroalcoholic, the alcohol content of these extraction steps is equal to or different from the initial solvent. In another particular embodiment, any of the above methods comprises a drying step, drying the extract.

[0039] In certain embodiments, a plant extract of one or more products of O. europaea as described in any aspect of the present invention is obtained by a method of obtaining a plant extract using a solvent as described in any of the previous embodiments, wherein the solvent is aqueous, preferably water.

[0040] In another particular embodiment, the plant extract of one or more products of Prunus dulcis as described in any aspect of the present invention is obtained by a method for obtaining a plant extract using a solvent as described in any of the previous embodiments, wherein the solvent is aqueous, preferably water.

[0041] In another specific embodiment, the plant extract of one or more products of Citrus paradisi as described in any aspect of the present invention is obtained by a method of obtaining a plant extract using a solvent as described in any of the previous embodiments, wherein the solvent is hydroalcoholic.

[0042] In another particular embodiment, the plant extract of one or more products of Linum usitatissimum as described in any aspect of the present invention is obtained by a method of obtaining a plant extract using a solvent as described in any of the previous embodiments, wherein the solvent is hydroalcoholic.

[0043] In another particular embodiment, the plant extract of one or more products of Polygonum cuspidatum as described in any of the aspects of the present invention is obtained by a method of obtaining a plant extract using a solvent as described in any of the previous embodiments, wherein the solvent is hydroalcoholic.

[0044] The term "activated charcoal" or "activated carbon" as used herein refers to a term known to those skilled in the art. It is a form of carbon that has been engineered to have increased adsorptive properties. Activated carbon is often engineered to have small pores with a low volume, which increases the surface area available for adsorption or chemical reaction.

[0045] As used herein, the term "product of a plant organism" or "plant product" refers to any part of a plant organism, including tissue of the plant organism, reproductive tissue of the plant organism, non-reproductive tissue of the plant organism, leaves, stems, roots, fruit, fruit tissue, fruit exocarp, fruit mesocarp, fruit endocarp, fruit pericarp, fruit shell, fruit pod, fruit seed, or pod of the plant organism. In certain embodiments, the plant product refers to the entirety of any of the parts of the plant organism just described. In another particular embodiment, the term refers to a portion of any of the parts of the plant organism just described.

[0046] Methods for determining the hydroxytyrosol content in plant extracts or the polyphenol content in plant extracts are generally known to those skilled in the art. These methods include any method that allows the determination of the amount and type of compounds in a composition, such as mass spectrometry. Preferred methods include mass spectrometry, high performance liquid chromatography-mass spectrometry (HPLC-MS), high performance liquid chromatography (HPLC)-ultraviolet absorption detector, gas chromatography-mass spectrometry (GC-MS), liquid chromatography-mass spectrometry (LC-MS), direct infusion mass spectrometry or Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR-MS), capillary electrophoresis-mass spectrometry (CE-MS), ultra-high performance liquid chromatography-mass spectrometry. (UHPLC-MS), supercritical fluid chromatography coupled to mass spectrometry (SFC-MS), flow injection analysis coupled to mass spectrometry (FIA-MS) (including quadrupole mass spectrometry), any sequential mass spectrometry such as MS-MS or MS-MS-MS, inductively coupled plasma mass spectrometry (ICPMS), pyrolysis-mass spectrometry (Py-MS), ion mobility mass spectrometry or time-of-flight (TOF) mass spectrometry, electrospray ionization mass spectrometry (ESIMS), ESI-MS / MS, ESI-(MS) <n>, matrix-assisted laser desorption / ionization time-of-flight mass spectrometry (MALDI-TOF-MS), surface-enhanced laser desorption / ionization time-of-flight mass spectrometry (SELDI-TOFMS), desorption / ionization on silicon (DIOS), secondary ion mass spectrometry (SIMS), quadrupole time-of-flight (Q-TOF), atmospheric pressure chemical ionization mass spectrometry (APCI-MS), APCI-MS / MS, APCI-(MS) <n>, Atmospheric Pressure Photoionization Mass Spectrometry (APPI-MS), APPI-MS / MS and APPI-(MS) <n>, quadrupole mass spectrometry, Fourier transform mass spectrometry (FTMS) and ion trap mass spectrometry, where n is an integer greater than 0. Such techniques are disclosed, for example, in Nissen, Journal of Chromatography A, 703, 1995: 37-57, U.S. Pat. No. 4,540,884 or U.S. Pat. No. 5,397,894. Preferably, gas chromatography coupled mass spectrometry is used as described in (Garcia A. and Barbas C. 2011, Methods Mol. Biol. (Clifton NJ) 708:191-204). Even more preferably, quadrupole time-of-flight gas chromatography coupled to mass spectrometry (GC-qTOF / MS) is used as described in (Riera-Borrull M. et al. 2016, J. Am. Soc. Mass Spectrom. 27(1):168-177; Kind T. et al., 2009, Anal. Chem. 81(24): 10038-48). In a preferred embodiment, the determination of the hydroxytyrosol level in the sample is performed by any of the above chromatographic techniques using a calibration line obtained with a hydroxytyrosol pattern of known concentration.

[0047] In a preferred embodiment, the determination of the polyphenol content in the plant extract is carried out using an enzymatic method, more preferably using a method based on Folin-Ciocalteu reagent (Singleton VL et al., 1999, Methods in enzimology, vol. 299: 152-78).

[0048] As used in the present invention, the expression "bioavailability" refers to the rate and speed at which an administered dose of a compound reaches its therapeutic target (cell receptor, channel, carrier), and reaching the therapeutic target means reaching the tissue where the compound acts. This is considered to be equivalent to the level reached in the systemic circulation of the subject who has taken or administered the compound. For this reason, in practical terms, bioavailability is specified by the percentage of the compound that occurs in plasma after administration of the compound.

[0049] As used in the present invention, the expression "bioavailability of hydroxytyrosol" refers to the bioavailability as defined above, in which the compound is hydroxytyrosol. Thus, the expression "bioavailability of hydroxytyrosol after oral administration of hydroxytyrosol" refers to the percentage of hydroxytyrosol in the systemic circulation, i.e. in the plasma, after oral administration of hydroxytyrosol to a subject. As a person skilled in the art will understand, the more stable hydroxytyrosol is in the digestive tract, the more hydroxytyrosol is available for absorption in the intestinal lumen, and the more hydroxytyrosol reaches the blood supply. Thus, the more stable hydroxytyrosol is in the digestive tract after oral administration of hydroxytyrosol, the higher its bioavailability. Thus, a method that allows for determining the bioavailability of hydroxytyrosol after oral administration of hydroxytyrosol includes determining the percentage of hydroxytyrosol in the serum of a subject administered with hydroxytyrosol, or determining the stability of hydroxytyrosol in the digestive tract of a subject administered with hydroxytyrosol orally. By way of example and not limitation, a method for determining the percentage of hydroxytyrosol in the serum of a subject can be any of the methods for determining the presence of polyphenols in a plant extract as indicated above, which are applied to a serum sample of the subject. By way of example and not limitation, a method that allows for determining the stability of hydroxytyrosol in the digestive tract can be the method described in Section B of Materials and Methods and the method described in Example 3 of the present application. The method can also be a modified version of the one described in that section of the present application, in particular the step of detecting the content of hydroxytyrosol in each of the compositions of the method of the present application can consist of any of the methods for detecting polyphenols in a plant extract as indicated above (in this case the polyphenol is hydroxytyrosol and the plant extract is replaced by the corresponding composition in which hydroxytyrosol was incubated in the method of the present application).

[0050] As used herein, the expression "increase the bioavailability of hydroxytyrosol after oral administration of hydroxytyrosol" refers to increase the bioavailability of hydroxytyrosol in a subject by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 110%, at least 120%, at least 130%, at least 140%, at least 150%, at least 160%, at least 170%, at least 180%, at least 190%, at least 200% based on a reference value. In a particular embodiment, the reference value is the bioavailability of hydroxytyrosol after oral administration of hydroxytyrosol without the combination of a polyphenol that is not hydroxytyrosol and does not naturally occur in O. europaea trees, and the polyphenol is as defined and described above and in different embodiments of the present invention. As will be understood by those skilled in the art, the reference value is the bioavailability of hydroxytyrosol after oral administration of a composition in which the only active ingredient is hydroxytyrosol. The composition may consist of a composition consisting of hydroxytyrosol and at least one pharma- ceutically acceptable excipient, which may be any of those indicated in the embodiments of the present invention for the pharmaceutical composition of the present invention. Methods for determining the bioavailability of hydroxytyrosol, such as those described immediately above, allow for determining the percentage by which the bioavailability of hydroxytyrosol is higher or lower relative to the reference value. As will be understood by those skilled in the art, the methods also allow for determining the bioavailability of hydroxytyrosol, which is defined as a reference value, and thus the reference value indicated in this definition.

[0051] The terms "atherosclerosis", "atherosclerotic vascular disease", or "ASVD" refer to cardiovascular disease characterized by chronic inflammation, lipid accumulation, alteration of vascular cells in the arterial wall, and the formation of atherosclerotic plaques or atheromas. The process leads to narrowing of the arterial lumen. Oxidized low-density lipoprotein (oxLDL) is considered to play a key role in the initiation and progression of atherogenesis. Because oxLDL binds with higher affinity to multiple receptors than native LDL, its absorption by macrophages is faster, which leads to cholesterol accumulation and the formation of atherosclerotic plaques or atheromas. Furthermore, oxLDL promotes endothelial cell activation and dysfunction, vascular smooth muscle cell migration and proliferation, and platelet activation, causing inflammatory responses, endothelial dysfunction, and exacerbating the atherogenic process. As a result, the concentration of circulating oxLDL is considered one of the most important biomarkers of atherosclerosis progression.

[0052] As used herein, the term "anti-atherosclerotic effect of hydroxytyrosol" refers to the activity of hydroxytyrosol once administered to a subject to help prevent, heal, reverse or treat atherosclerosis. Suitable indicators of the anti-atherosclerotic effect of hydroxytyrosol include a reduction in serum oxLDL concentration or a reduction in serum oxLDL / LDL-cholesterol ratio after administration, preferably orally, to a subject.

[0053] As used in the present invention, the expression "LDL", "LDL-c" or "LDL-cholesterol" refers to the term that is generally known to those skilled in the art. This term specifically refers to low density lipoprotein, i.e. LDL. LDL is one of the five main groups of lipoproteins (ULD, very low density lipoprotein; VLDL, very low density lipoprotein; IDL, intermediate density lipoprotein; HDL, high density lipoprotein; and LDL as defined herein) that transport fat molecules in the extracellular medium in the body. Most cholesterol is transported in the subject's blood together with protein to form LDL.

[0054] As used in the present invention, the term "ox-LDL" or "oxLDL" refers to oxidized LDL lipoproteins as defined above. Oxidized LDL is a general term for LDL lipoproteins with structural elements modified by oxidation. As a result of attack by free radicals, both the lipid and protein parts of LDL can be oxidized. In addition to oxidation reactions occurring in the vascular wall giving rise to ox-LDL, oxidation of lipids in LDL can also result from dietary oxidized lipids. ox-LDL is associated with the development of atherosclerosis. The effect of oxidized LDL on atherogenesis has been explained by the failure of LDL receptors to recognize the structure of LDL modified by oxidation, which prevents the normal metabolism of LDL particles from occurring, ultimately leading to the development of atherosclerotic plaques.

[0055] As will be appreciated by those skilled in the art, the expressions "ox-LDL / LDL-cholesterol" or "ox-LDL / LDL" as used in the present invention refer to the ratio between the concentration in a particular ox-LDL medium, as defined above, and the LDL concentration in said medium, as defined above.

[0056] As used herein, the expression "increasing the anti-atherosclerotic effect of hydroxytyrosol" refers to increasing any of the effects set out in the above specific embodiments in the definition of atherosclerotic effect by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 110%, at least 120%, at least 130%, at least 140%, at least 150%, at least 160%, at least 170%, at least 180%, at least 190%, at least 200%, at least 210%, at least 220%, at least 230%, at least 240%, at least 250%, at least 275%, at least 300%, at least 350%, at least 400% relative to a reference value. In a particular embodiment, the reference value is the value of the parameter change due to administration of hydroxytyrosol as indicated in the corresponding embodiment above in the definition of the atherosclerotic effect upon administration of hydroxytyrosol without a polyphenol other than hydroxytyrosol and not naturally occurring in the O. europaea tree, said polyphenol being as defined and described above and in the different embodiments of the invention. Preferably, the hydroxytyrosol administered to determine the reference value consists of a composition in which the active compound is hydroxytyrosol only, in particular a composition comprising only hydroxytyrosol and optionally a pharma- ceutically acceptable excipient, the excipient being as defined in the aspects of the invention for the pharmaceutical composition of the invention.

[0057] In a particular embodiment, the expression "increase the anti-atherosclerotic effect of hydroxytyrosol" refers to reducing serum oxLDL concentration by at least 10%, at least 20%, at least 30%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 110%, at least 120%, at least 130%, at least 140%, at least 150%, at least 160%, at least 170%, at least 180%, at least 190%, at least 200%, at least 210%, at least 220%, at least 230%, at least 240%, at least 250%, at least 275%, at least 300%, at least 350%, at least 400%, preferably at least 20%, based on a reference value. In a particular embodiment, this reduction based on a reference value is at least 50%. In another particular embodiment, this is at least 70%. In a particular embodiment, the reference value is the serum oxLDL concentration after oral administration of hydroxytyrosol without concomitant polyphenols that are not hydroxytyrosol and do not naturally occur in the O. europaea tree, the polyphenols being as defined and described above and in the different embodiments of the present invention. Preferably, the hydroxytyrosol administered to determine the reference value consists of a composition in which the active compound is hydroxytyrosol alone, in particular a composition comprising only hydroxytyrosol and optionally a pharma-ceutically acceptable excipient, the excipient being as defined in the aspect of the present invention for the pharmaceutical composition of the present invention. The method of determining the reduction comprises the method described in Section A of the Materials and Methods of the present application (in particular in the "Analysis of Biochemical Parameters" section) for determining the serum oxLDL concentration and comparing the obtained value with the reference value.In another particular embodiment, the expression "increase the anti-atherosclerotic effect of hydroxytyrosol" refers to reduce the serum oxLDL / LDL-cholesterol ratio by at least 10%, at least 20%, at least 30%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 70%, at least 80%, at least 90%, at least 100%, at least 110%, at least 120%, at least 130%, at least 140%, at least 150%, at least 160%, at least 170%, at least 180%, at least 190%, at least 200%, at least 210%, at least 220%, at least 230%, at least 240%, at least 250%, at least 275%, at least 300%, at least 350%, at least 400%, preferably at least 25%, based on the reference value.In a particular embodiment, this reduction is at least 30% based on the reference value. In another particular embodiment, this is at least 50%. In a particular embodiment, the reference value is the serum oxLDL / LDL-cholesterol ratio after oral administration of hydroxytyrosol without concomitant polyphenols that are not hydroxytyrosol and do not naturally occur in the O. europaea tree, the polyphenols being as defined and described above and in the different embodiments of the invention. Preferably, the hydroxytyrosol administered to determine the reference value consists of a composition in which the active compound is only hydroxytyrosol, in particular a composition comprising only hydroxytyrosol and optionally a pharma-ceutically acceptable excipient, the excipient being as defined in the aspect of the invention for the pharmaceutical composition of the invention. Methods for determining the reduction include the methods for determining serum oxLDL concentrations described in Section A of the Materials and Methods of the present application (in particular in the "Analysis of Biochemical Parameters" section) and the methods for determining serum LDL concentrations generally known to those skilled in the art.Non-limiting examples of such methods include the direct method (GD Garcia Aguilar et al., 2006, Quimica Clinica, 25(2): 58-63), the Friedman method (Friedewald WT et al., Clin Chem. 1972;18(6):499-502) or the beta quantification method using ultracentrifugation (Warnick GR et al. Lab Med. 2008;39(8):481-90). The ratio between the values ​​is then determined and the value(s) of the oxLDL / LDL-cholesterol ratio obtained in the serum sample of interest is compared with the reference value.

[0058] In certain embodiments of the first, second and third aspects of the invention, the at least one polyphenol that is not hydroxytyrosol and / or that does not naturally occur in Olea europaea is administered in combination with hydroxytyrosol, i.e. as part of the same composition. Alternatively, in certain embodiments of the first, second and third aspects of the invention, the at least one polyphenol that is not hydroxytyrosol itself and / or that does not naturally occur in Olea europaea is administered sequentially with respect to hydroxytyrosol. Alternatively, in certain embodiments of the first, second and third aspects of the invention, the at least one polyphenol that is not hydroxytyrosol itself and / or that does not naturally occur in Olea europaea is administered separately with respect to hydroxytyrosol.

[0059] In certain embodiments of the first, second and third aspects of the invention, the hydroxytyrosol is part of a plant extract.

[0060] The expression "plant extract" is defined above. In a particular embodiment, the plant extract containing hydroxytyrosol is obtained from any plant product of the O. europaea tree, which plant product is any of those indicated in the definition of "plant product" given above. In a preferred embodiment of the first, second and third aspects of the invention, the plant extract of which hydroxytyrosol is a part is an extract of the leaves or fruits of the O. europaea tree, preferably the leaves.

[0061] As used herein, the phrase "is a part of a plant extract" indicates that the particular compound is contained in the plant extract, and the terms "comprised" and "comprise" indicate that the plant extract must contain the compound in question, but may optionally contain further compounds or ingredients.

[0062] In a particular embodiment, the plant extract comprising hydroxytyrosol is obtained from at least two plant products of the O. europaea tree, the plant products being selected from those set out in the definition of the plant product, each of said products being the same or different, in another particular embodiment, each of said products is obtained from a different tree of O. europaea, each tree being preferably of a different subspecies, more preferably each tree being of a different variety.

[0063] In a particular embodiment, the tree of the O. europaea species referred to in any definition or in a particular embodiment of any aspect of the invention is any variety of the tree of the O. europaea species. In another particular embodiment, it is a wild variety of the common olive tree, i.e., a variety of Olea europaea sylvestris. In another particular embodiment, it is a tree of a subspecies selected from the group consisting of: O. europaea europaea, O. europaea kerasiformis, O. europaea cuspidata, O. europaea guantica, O. europaea lapellinii, and O. europaea maroccana. In another particular embodiment, it is any variety of the above subspecies.

[0064] In a particular embodiment, the O. europaea tree referred to in any definition or in any particular embodiment of any aspect of the invention is each of any variety of O. europaea tree. In another particular embodiment, at least one is a wild variety of the common olive tree, i.e., a variety of Olea europaea sylvestris. In another embodiment, at least one is a tree of a subspecies selected from the group consisting of: O. europaea subspecies europaea, O. europaea kerasiformis, O. europaea cuspidata, O. europaea guantica, O. europaea lapellinii, and O. europaea maroccana. In another particular embodiment, at least one is a variety of any of the above subspecies.

[0065] In certain embodiments, the plant extract containing hydroxytyrosol contains hydroxytyrosol at 1% to 100% (w / w), 1% to 98% (w / w), 1% to 95% (w / w), 1% to 90% (w / w), 3% to 90% (w / w), 5% to 90% (w / w), 7% to 90% (w / w), 8% to 90% (w / w), 9% to 90%(w / w), 10%~90%(w / w), 10%~85%(w / w), 10%~80%(w / w), 10%~70%(w / w), 10%~60%(w / w), 10% to 50% (w / w), 10% to 40% (w / w), 10% to 30% (w / w), 10% to 25% (w / w), 10% to 20% (w / w). In a preferred embodiment, the plant extract containing hydroxytyrosol contains hydroxytyrosol at 1% (w / w), 2% (w / w), 3% (w / w), 4% (w / w), 5% (w / w), 6% (w / w), 7% (w / w), 8% (w / w), 9% (w / w), 10% (w / w), 11% (w / w), 12% (w / w), 13% (w / w), 14% (w / w), 15% (w / w), 16% (w / w), 17% (w / w), 18% (w / w), 19% (w / w), 20% (w / w), 22% (w / w), 25% (w / w), 27% (w / w) , 30%(w / w), 35%(w / w), 40%(w / w), 50%(w / w), 55%(w / w), 60%(w / w), 65%(w / w), 70%(w / w), 75%(w / w), 80%(w / w), 90%(w / w), 95%(w / w), 97%(w / w), 98% (w / w), 99%(w / w), 99.25%(w / w), 99.5%(w / w), 99.7%(w / w), 99.8%(w / w), 9 It contains 9.9% (w / w), 99.95% (w / w), 9.99% (w / w), 100% (w / w), preferably 10% (w / w).

[0066] In certain embodiments of the first, second and third aspects of the present invention, the hydroxytyrosol-containing plant extract contains 10% to 90% hydroxytyrosol, preferably 10% (w / w).

[0067] In another particular embodiment of the first, second and third aspects of the invention, the at least one polyphenol is selected from the group of polyphenols consisting of flavonoids, lignans and stilbenes.

[0068] As used herein, the terms "flavonoids," "lignans," and "stilbenes" refer to polyphenols commonly known by that name to those skilled in the art. Specifically, they refer to polyphenols as defined above.

[0069] In certain embodiments of the first, second and third aspects of the present invention, the at least one polyphenol other than hydroxytyrosol in the group of flavonoids is selected from the group consisting of procyanidins, propelargonidins, prodelphinidins, catechins, epicatechins, quercetins, kaempferol, isorhamnetin, naringin, naringenin, genistein and rutin.

[0070] As used herein, the terms "procyanidin," "propelargonidin," "prodelphinidin," "catechin," "epicatechin," "quercetin," "kaempferol," "isorhamnetin," "naringin," "naringenin," "genistein," and "rutin" refer to the corresponding terms as understood by those of skill in the art.

[0071] As used herein, the term "procyanidin" refers to a group of proanthocyanidins that are composed only of (epi)catechin. As used herein, the term "propelargonidin" refers to a proanthocyanidin that has an (epi)afzeletin unit in combination with an (epi)catechin unit. As used herein, the term "prodelphinidin" refers to a proanthocyanidin that has at least one (epi)gallocatechin unit in combination with an (epi)catechin unit. As used herein, the term "proanthocyanidin" or "condensed tannin" refers to oligomers and polymers of flavan-3-ols that are widely distributed in the plant kingdom. The most abundant flavan-3-ol units are (+)-afzeletin, (+)-catechin, and (+)-gallocatechin (2R:3S configuration) and their respective diastereoisomers, (-)-epiafzeletin, (-)-epicatechin, and (-)-epigallocatechin (2R:3R configuration). In the B-type procyanidins / propelargonidins / prodelphinidins, the flavan-3-ol units are linked at the C-4 carbon of the upper unit to the C-6 or C-8 carbon of the lower unit, with the C4-C8 isomers being more prevalent than the C4-C6 isomers. In addition to this interflavan C-C bond, A-type procyanidins have an ether-type bond between the C-2 carbon of the upper unit and the hydroxyl group of the C-7 carbon of the lower unit (Porter LJ (1988) Flavans and proanthocyanidins. In The flavonoids (Harborne JB, Ed.) Chapman and Hall, New York, pp. 21-62).

[0072] Therefore, in certain embodiments, the polyphenol other than hydroxytyrosol in the flavonoid group shown in any aspect of the present invention can be any polyphenol belonging to the proanthocyanidin group.In certain embodiments, it is any polyphenol in the procyanidin group, and preferably it is B-type procyanidin.In certain embodiments, it is a procyanidin selected from the group consisting of B1-type procyanidin, B2-type procyanidin, B3-type procyanidin, B4-type procyanidin, B5-type procyanidin, B6-type procyanidin, and B7-type procyanidin.In another specific embodiment, it is a procyanidin of C1 type.In a specific embodiment, it is a procyanidin of A type, and preferably it is selected from the group consisting of procyanidin A1 and procyanidin A2.In another specific embodiment, it is any polyphenol in the propelargonidin group, and preferably it is A-type propelargonidin. In another particular embodiment, it is type A propelargonidin, more preferably type B propelargonidin. In another particular embodiment, it is any polyphenol of the prodelphinidin group, and in a particular embodiment, preferably type A prodelphinidin.

[0073] Specifically, as used herein, the term "procyanidin B1" or "B1-type procyanidin" refers to the compound having the IUPAC name of (2R,3S)-2-(3,4-dihydroxyphenyl)-8-[(2R,3R,4R)-2-(3,4-dihydroxyphenyl)-3,5,7-trihydroxy-3,4-dihydro-2H-chromen-4-yl]-3,4-dihydro-2H-chromene-3,5,7-triol.

[0074] As used herein, the term "procyanidin B2" or "B2-type procyanidin" refers to the compound having the IUPAC name (2R,3R)-2-(3,4-dihydroxyphenyl)-8-[(2R,3R,4R)-2-(3,4-dihydroxyphenyl)-3,5,7-trihydroxy-3,4-dihydro-2H-chromen-4-yl]-3,4-dihydro-2H-chromene-3,5,7-triol.

[0075] As used herein, the term "procyanidin B3" or "B3-type procyanidin" refers to the compound having the IUPAC name (2R,3S)-2-(3,4-dihydroxyphenyl)-8-[(2R,3S,4S)-2-(3,4-dihydroxyphenyl)-3,5,7-trihydroxy-3,4-dihydro-2H-chromen-4-yl]-3,4-dihydro-2H-chromene-3,5,7-triol.

[0076] As used herein, the term "procyanidin B4" or "B3-type procyanidin" refers to the compound having the IUPAC name (2R,3R)-2-(3,4-dihydroxyphenyl)-8-[(2R,3S,4S)-2-(3,4-dihydroxyphenyl)-3,5,7-trihydroxy-3,4-dihydro-2H-chromen-4-yl]-3,4-dihydro-2H-chromene-3,5,7-triol.

[0077] As used herein, the term "procyanidin B5" or "B5-type procyanidin" refers to the compound having the IUPAC name (2R,3R)-2-(3,4-dihydroxyphenyl)-6-[(2R,3R,4S)-2-(3,4-dihydroxyphenyl)-3,5,7-trihydroxy-3,4-dihydro-2H-chromen-4-yl]-3,4-dihydro-2H-chromene-3,5,7-triol.

[0078] As used herein, the term "procyanidin B6" or "B6-type procyanidin" refers to the compound having the IUPAC name (2R,3S)-2-(3,4-dihydroxyphenyl)-6-[(2R,3S,4R)-2-(3,4-dihydroxyphenyl)-3,5,7-trihydroxy-3,4-dihydro-2H-chromen-4-yl]-3,4-dihydro-2H-chromene-3,5,7-triol.

[0079] The term "procyanidin B7" or "procyanidin type B7" as used herein refers to the compound named (2R,3S)-2-(3,4-dihydroxyphenyl)-6-[(2R,3R,4S)-2-(3,4-dihydroxyphenyl)-3,5,7-trihydroxy-3,4-dihydro-2H-chromen-4-yl]-3,4-dihydro-2H-chromen-3,5,7-triol.

[0080] As used herein, the term "procyanidin C1" or "C1-type procyanidin" refers to the compound having the IUPAC name (2R,3R,4S)-2-(3,4-dihydroxyphenyl)-4-[(2R,3R)-2-(3,4-dihydroxyphenyl)-3,5,7-trihydroxy-3,4-dihydro-2H-chromen-8-yl]-8-[(2R,3R,4R)-2-(3,4-dihydroxyphenyl)-3,5,7-trihydroxy-3,4-dihydro-2H-chromen-4-yl]-3,4-dihydro-2H-chromene-3,5,7-triol.

[0081] As used herein, the term "procyanidin A1" or "A1-type procyanidin" refers to the compound having the IUPAC name (1R,5R,6S,13S,21R)-5,13-bis(3,4-dihydroxyphenyl)-4,12,14-trioxapentacyclo[11.7.1.02,11.03,8.015,20]heneicosa-2(11),3(8),9,15,17,19-hexaene-6,9,17,19,21-pentaol.

[0082] As used herein, the term "procyanidin A2" or "A2-type procyanidin" refers to the compound having the IUPAC name (1R,5R,6R,13S,21R)-5,13-bis(3,4-dihydroxyphenyl)-4,12,14-trioxapentacyclo[11.7.1.02,11.03,8.015,20]heneicosa-2(11),3(8),9,15,17,19-hexaene-6,9,17,19,21-pentaol.

[0083] As used herein, the term "propelargonidin" refers to the compound having the IUPAC name 2-[[2-(3,4-dihydroxyphenyl)-5,7-dihydroxy-3,4-dihydro-2H-chromen-3-yl]oxy]-2-(4-hydroxyphenyl)-3,4-dihydrochromene-3,4,5,7-tetraol.

[0084] In another particular embodiment of the first, second and third aspects of the present invention, the at least one polyphenol from the lignan group other than hydroxytyrosol is selected from the group consisting of secoisolariciresinol diglucoside, secoisolariciresinol, matairesinoside, lariciresinol, pinoresinol, sesamin and secoisolariciresinol.

[0085] As used herein, the terms "secoisolariciresinol diglucoside," "matairesinoside," "lariciresinol," "pinoresinol," "sesamin," and "secoisolariciresinol" refer to the corresponding terms as understood by those of skill in the art.

[0086] In another particular embodiment of the first, second and third aspects of the present invention, the at least one polyphenol from the stilbenoid group other than hydroxytyrosol is selected from the group consisting of resveratrol, piceatannol, pinosylvin, pterostilbene and rhapontigenin.

[0087] As used herein, the terms "resveratrol", "piceatannol", "pinosylvin", "pterostilbene", and "rhapontigenin" refer to the corresponding terms as understood by those of skill in the art.

[0088] In certain embodiments of the first, second and third aspects of the invention, the at least one polyphenol other than hydroxytyrosol is part of a plant extract.

[0089] In another particular embodiment, the plant extract is not an extract of a plant product of the O. europaea tree, and preferably it is not an extract of the leaves or fruits of the O. europaea tree.

[0090] In another particular embodiment of the first, second and third aspect of the invention, the plant extract of which at least one polyphenol other than hydroxytyrosol is a part is an extract selected from the group consisting of: Extract of plant products from Prunus dulcis (Miller) DA Webb Citrus Paradisi MacFad Plant Product Extract Extract of plant products from Linum usitatissimum L. Extract of plant products from Polygonum cuspidatum Sieb. et Zucc

[0091] In certain embodiments, each of the plant products referred to in the foregoing embodiments is one of those set forth in the definition of "plant product" provided above.

[0092] In a preferred embodiment, the plant extract of which at least one polyphenol other than hydroxytyrosol is a part is an extract selected from the group consisting of: Extract of the peel of Prunus dulcis (Miller) DA Webb Citrus paradisi MacFad fruit extract or Citrus paradisi MacFad fruit seed extract Extract of the fruit seeds of Linum usitatissimum L. Polygonum cuspidatum Sieb. et Zucc root extract

[0093] The plant extracts of which at least one polyphenol other than hydroxytyrosol is a part, as specified in the above embodiments, can be obtained from any plant or vegetable belonging to the species indicated in each example, preferably any variety belonging to the species indicated in each example. In a particular embodiment, the tree is of the species Prunus dulcis (Miller) DA Webb.

[0094] In a particular embodiment, the term Prunus dulcis used in any aspect of the present invention is synonymous with the term Prunus dulcis (Miller) DA Webb. In another particular embodiment, the term Citrus paradisi used in any aspect of the present invention is synonymous with the term Citrus paradisi MacFad. In another particular embodiment, the term Linum usitatissimum used in any aspect of the present invention is synonymous with the term Linum usitatissimum L. In another particular embodiment, the term Polygonum cuspidatum used in any aspect of the present invention is synonymous with the term Polygonum cuspidatum Sieb.et Zucc.

[0095] In a particular embodiment, the plant extract containing polyphenols other than hydroxytyrosol obtained from a particular species is obtained from two or more plant products of a tree, plant or vegetable of that species, each of which may be the same or different.In another particular embodiment, each of the products is obtained from a different tree, plant or vegetable, each of which may be of the same species, preferably of the same variety.

[0096] Thus, in a particular embodiment, the plant extract of which at least one polyphenol other than hydroxytyrosol is a part is an extract selected from the group consisting of: Extracts of at least two plant products, each selected from the list of plant products set out in the definition of "Plant Product" above, are obtained from at least two different Prunus dulcis trees, each tree preferably being of a different variety. Extracts of at least two plant products, each selected from the list of plant products set out in the definition of "Plant Product" above, are obtained from at least two different Citrus Paradisi trees, each tree preferably being of a different variety. Extracts of at least two plant products, each selected from the list of plant products set out in the definition of "Plant Product" above, are obtained from at least two different Linum usitatissimum plants, each plant preferably being of a different variety. Extracts of at least two plant products, each selected from the list of plant products set out in the definition of "Plant Product" above, are obtained from at least two different Polygonum cuspidatum plants, each plant preferably being of a different variety.

[0097] In certain embodiments, The at least one polyphenol other than hydroxytyrosol that is part of any of the above extracts of one or more plant products of one or more trees of Prunus, preferably an extract of the pericarp, is a flavonoid, preferably a flavonoid selected from the group consisting of procyanidin, propelargonidin, prodelphinidin, catechin, epicatechin, quercetin, kaempferol, isorhamnetin, rutin, and genistein. At least one polyphenol other than hydroxytyrosol that is part of any of the above extracts of one or more plant products of one or more trees of Citrus paradisi, preferably an extract of the seeds of the fruit, is a flavonoid, preferably a flavonoid selected from the group consisting of naringin, naringenin, quercetin, catechin, rutin, and genistein. At least one polyphenol other than hydroxytyrosol that is part of any of the above extracts of one or more plant products of Linum usitatissimum, preferably an extract of the fruit seeds, is a lignan, preferably a lignan selected from the group consisting of secoisolariciresinol diglucoside, secoisolariciresinol, matairesinoside, lariciresinol, pinoresinol, and sesamin. At least one polyphenol other than hydroxytyrosol that is part of any of the above extracts, preferably root extracts, of one or more plant products of one or more plants of Polygonum cuspidatum is a stilbene, preferably a stilbene selected from the group consisting of resveratrol, piceatannol, pinosylvin, pterostilbene, and rhapontigenin.

[0098] In preferred embodiments of the first, second and third aspects of the present invention, The at least one polyphenol other than hydroxytyrosol that is part of the extract of the pericarp of Prunus dulcis is a flavonoid, preferably selected from the group consisting of procyanidin, propelargonidin, prodelphinidin, catechin, epicatechin, quercetin, kaempferol, isorhamnetin, rutin, and genistein. At least one polyphenol other than hydroxytyrosol that is part of the Citrus Paradisi fruit extract or Citrus Paradisi fruit seed extract is a flavonoid, preferably selected from the group consisting of naringin, naringenin, quercetin, catechin, rutin, and genistein. The at least one polyphenol other than hydroxytyrosol which is part of the extract of the seeds of the fruit of Linum usitatissimum is a lignan, preferably selected from the group consisting of secoisolariciresinol diglucoside, secoisolariciresinol, matairesinoside, lariciresinol, pinoresinol, and sesamin.

[0099] In a particular embodiment, any of the above extracts of one or more plant products of one or more trees of Prunus dulcis, preferably an extract of the pericarp, contains 1% to 100% (w / w), 1% to 90% (w / w), 5% to 80% (w / w), 5% to 70% (w / w), 5% to 80% (w / w), 5% to 60% (w / w), 5% to 50% (w / w), 5% to 40% (w / w), 10% to 30% (w / w), preferably 5% to 60% (w / w) of flavonoids. Preferably, the flavonoids are selected from the group consisting of procyanidins, propelargonidins, prodelphinidins, catechins, epicatechins, quercetin, kaempferol, isorhamnetin, rutin, genistein, and combinations thereof. Particular embodiments include those comprising flavonoids at 1% (w / w), 2% (w / w), 3% (w / w), 4% (w / w), 5% (w / w), 6% (w / w), 7% (w / w), 8% (w / w), 9% (w / w), 10% (w / w), 12% (w / w), 15% (w / w), 20% (w / w), 22% (w / w), 25% (w / w), 27% (w / w), 28% (w / w), 29% (w / w), 30% (w / w), 31% (w / w), 32% (w / w), 33% (w / w), 34% (w / w), 35% (w / w), 36% (w / w), 37% (w / w), 38% (w / w), 39% (w / w), 40% (w / w), 41% (w / w), 42% (w / w), 43% (w / w), 44% (w / w), 45% (w / w), 46% (w / w), 47% (w / w), 48% (w / w), 49% (w / w), 50% (w / w), 51% (w / w), 52% (w / w), 53% (w / w), 54% (w / w), 55% (w / w), 56% (w / w), 57% (w / w), 58% (w / w), 59% (w / w), 60% (w / w), 61% (w / w), 62% (w / w), 63% (w / w), 64% (w / w), 65% (w / w), 66% (w / w), 67% (w / w), 68% %(w / w), 32%(w / w), 35%(w / w), 40%(w / w), 45%(w / w), 50%(w / w), 60%(w / w), 70%(w / w), 80%(w / w), 90%(w / w), 95%(w / w), 97%(w / w), 98%(w / w), 99%(w / w), 99.5%(w / w), 99.8%(w / w), 99.9%(w / w), 100%(w / w), preferably 30%(w / w). Preferably, the flavonoid is selected from the list of flavonoids given above. In a preferred embodiment, the determination of the polyphenol content is carried out using a method based on the Folin-Ciocalteu reagent (Singleton VL et al., 1999, Methods in Enzymology, vol. 299: 152-78).

[0100] In a preferred embodiment, the extract of the pericarp of Prunus dulcis contains flavonoids at 1% to 100% (w / w), 1% to 90% (w / w), 5% to 80% (w / w), 5% to 70% (w / w), 5% to 80% (w / w), 5% to 60% (w / w), 5% to 50% (w / w), 5% to 40% (w / w), 10% to 30% (w / w), preferably 5% to 60% (w / w). Preferably, the flavonoids are selected from the group consisting of procyanidin, propelargonidin, prodelphinidin, catechin, epicatechin, quercetin, kaempferol, isorhamnetin, rutin, genistein, and combinations thereof. In certain embodiments, the extract contains flavonoids at 1% (w / w), 2% (w / w), 3% (w / w), 4% (w / w), 5% (w / w), 6% (w / w), 7% (w / w), 8% (w / w), 9% (w / w), 10% (w / w), 12% (w / w), 15% (w / w), 20% (w / w), 22% (w / w), 25% (w / w), 27% (w / w), 28% (w / w), 29% (w / w), 30% (w / w), 31% (w / w), 32% (w / w), 35% (w / w), 40% (w / w), 45% (w / w), 50% (w / w), 60% (w / w), 70% (w / w), 80% (w / w), 90% (w / w), 95% (w / w), 97% (w / w), 98% (w / w), 99% (w / w), 99.5% (w / w), 99.8% (w / w), 99.9% (w / w), 100% (w / w), preferably 30% (w / w). Preferably, the flavonoid is selected from the list of flavonoids given above. In a preferred embodiment, the determination of the polyphenol content is carried out using a method based on the Folin-Ciocalteu reagent (Singleton VL et al., 1999, Methods in Enzymology, vol. 299: 152-78).

[0101] In another particular embodiment, any of the above extracts of one or more plant products of one or more trees of Citrus paradisi, preferably fruit or fruit seed extracts, contains flavonoids in an amount of 1% to 100% (w / w), 1% to 90% (w / w), 5% to 80% (w / w), 5% to 70% (w / w), 5% to 80% (w / w), 10% to 80% (w / w), 10% to 60% (w / w), 15% to 50% (w / w), 20% to 50% (w / w), 30% to 50% (w / w), preferably 10% to 80% (w / w). Preferably, the flavonoids are selected from the list consisting of naringin, naringenin, quercetin, catechin, rutin, genistein, and combinations thereof. In certain embodiments, the extract contains flavonoids at 1% (w / w), 2% (w / w), 3% (w / w), 4% (w / w), 5% (w / w), 6% (w / w), 7% (w / w), 8% (w / w), 9% (w / w), 10% (w / w), 12% (w / w), 15% (w / w), 20% (w / w), 22% (w / w), 25% (w / w), 27% (w / w), 28% (w / w), 29% (w / w), 30% (w / w), 31% (w / w), 32% (w / w), 35% (w / w), 40% (w / w), 45% (w / w), 50% (w / w), 60% (w / w), 70% (w / w), 80% (w / w), 90% (w / w), 95% (w / w), 97% (w / w), 98% (w / w), 99% (w / w), 99.5% (w / w), 99.8% (w / w), 99.9% (w / w), 100% (w / w), preferably 45% (w / w). Preferably, the flavonoid is selected from the list consisting of naringin, naringenin, quercetin, catechin, rutin, genistein, and combinations thereof. In a preferred embodiment, the determination of the polyphenol content is carried out using a method based on the Folin-Ciocalteu reagent (Singleton VL et al., 1999, Methods in enzimology, vol. 299: 152-78).

[0102] In a preferred embodiment, the extract of the fruit of Citrus paradisi or the extract of the seeds of the fruit of Citrus paradisi contains flavonoids in an amount of 1% to 100% (w / w), 1% to 90% (w / w), 5% to 80% (w / w), 5% to 70% (w / w), 5% to 80% (w / w), 10% to 80% (w / w), 10% to 60% (w / w), 15% to 50% (w / w), 20% to 50% (w / w), 30% to 50% (w / w), preferably 10% to 80% (w / w). Preferably, the flavonoids are selected from the list consisting of naringin, naringenin, quercetin, catechin, rutin, genistein, and combinations thereof. In certain embodiments, the extract contains flavonoids at 1% (w / w), 2% (w / w), 3% (w / w), 4% (w / w), 5% (w / w), 6% (w / w), 7% (w / w), 8% (w / w), 9% (w / w), 10% (w / w), 12% (w / w), 15% (w / w), 20% (w / w), 22% (w / w), 25% (w / w), 27% (w / w), 28% (w / w), 29% (w / w), 30% (w / w), 31% (w / w), 32% (w / w), 35% (w / w), 40% (w / w), 45% (w / w), 50% (w / w), 60% (w / w), 70% (w / w), 80% (w / w), 90% (w / w), 95% (w / w), 97% (w / w), 98% (w / w), 99% (w / w), 99.5% (w / w), 99.8% (w / w), 99.9% (w / w), 100% (w / w), preferably 45% (w / w). Preferably, the flavonoid is selected from the list consisting of naringin, naringenin, quercetin, catechin, rutin, genistein, and combinations thereof. In a preferred embodiment, the determination of the polyphenol content is carried out using a method based on the Folin-Ciocalteu reagent (Singleton VL et al., 1999, Methods in Enzymology, vol. 299: 152-78).

[0103] In another particular embodiment, any of the above extracts of one or more plant products of one or more plants of Linum usitatissimum, preferably fruit seed extracts, contain lignans in an amount of 1%-100% (w / w), 1%-90% (w / w), 5%-80% (w / w), 5%-70% (w / w), 5%-80% (w / w), 5%-60% (w / w), 5%-50% (w / w), 5%-40% (w / w), 10%-30% (w / w), preferably 5%-50% (w / w) of lignans. Preferably, the lignans are selected from the group consisting of secoisolariciresinol diglucoside, secoisolariciresinol, matairesinoside, lariciresinol, pinoresinol, and sesamin, and combinations thereof. In certain embodiments, the extract contains lignans at 1% (w / w), 2% (w / w), 3% (w / w), 4% (w / w), 5% (w / w), 6% (w / w), 7% (w / w), 8% (w / w), 9% (w / w), 10% (w / w), 12% (w / w), 15% (w / w), 20% (w / w), 22% (w / w), 25% (w / w), 27% (w / w), 28% (w / w), 29% (w / w), 30% (w / w), 31% (w / w), 32% (w / w), 33% (w / w), 34% (w / w), 35% (w / w), 36% (w / w), 37% (w / w), 38% (w / w), 39% (w / w), 40% (w / w), 41% (w / w), 42% (w / w), 43% (w / w), 44% (w / w), 45% (w / w), 46% (w / w), 47% (w / w), 48% (w / w), 49% (w / w), 50% (w / w), 51% (w / w), 52% (w / w), 53% (w / w), 54% (w / w), 55% (w / w), 56% (w / w), 57% (w / w), 58% (w / w), 59% (w / w), 60% (w / w), 61% (w / w), 62% (w / w), 63% (w / w), 64% (w / w), 65% (w / w), 66% (w / w), 67% (w / w), 68% The lignans may be present in an amount of 1% (w / w), 32% (w / w), 35% (w / w), 40% (w / w), 45% (w / w), 50% (w / w), 60% (w / w), 70% (w / w), 80% (w / w), 90% (w / w), 95% (w / w), 97% (w / w), 98% (w / w), 99% (w / w), 99.5% (w / w), 99.8% (w / w), 99.9% (w / w), 100% (w / w), preferably 20% (w / w). Preferably, the lignans are selected from the above list. In a preferred embodiment, the determination of the polyphenol content is carried out using a method based on the Folin-Ciocalteu reagent (Singleton VL et al., 1999, Methods in enzimology, vol. 299: 152-78).

[0104] In a preferred embodiment, the extract of seeds of Linum usitatissimum contains lignans at 1%-100% (w / w), 1%-90% (w / w), 5%-80% (w / w), 5%-70% (w / w), 5%-80% (w / w), 5%-60% (w / w), 5%-50% (w / w), 5%-40% (w / w), 10%-30% (w / w), preferably 5%-50% (w / w) of lignans. Preferably, the lignans are selected from the group consisting of secoisolariciresinol diglucoside, secoisolariciresinol, matairesinoside, lariciresinol, pinoresinol, and sesamin, and combinations thereof. In certain embodiments, the extract contains lignans at 1% (w / w), 2% (w / w), 3% (w / w), 4% (w / w), 5% (w / w), 6% (w / w), 7% (w / w), 8% (w / w), 9% (w / w), 10% (w / w), 12% (w / w), 15% (w / w), 20% (w / w), 22% (w / w), 25% (w / w), 27% (w / w), 28% (w / w), 29% (w / w), 30% (w / w), 31% (w / w), 32% (w / w), 33% (w / w), 34% (w / w), 35% (w / w), 36% (w / w), 37% (w / w), 38% (w / w), 39% (w / w), 40% (w / w), 41% (w / w), 42% (w / w), 43% (w / w), 44% (w / w), 45% (w / w), 46% (w / w), 47% (w / w), 48% (w / w), 49% (w / w), 50% (w / w), 51% (w / w), 52% (w / w), 53% (w / w), 54% (w / w), 55% (w / w), 56% (w / w), 57% (w / w), 58% (w / w), 59% (w / w), 60% (w / w), 61% (w / w), 62% (w / w), 63% (w / w), 64% (w / w), 65% (w / w), 66% (w / w), 67% (w / w), 68% The lignans may be present in an amount of 1% (w / w), 32% (w / w), 35% (w / w), 40% (w / w), 45% (w / w), 50% (w / w), 60% (w / w), 70% (w / w), 80% (w / w), 90% (w / w), 95% (w / w), 97% (w / w), 98% (w / w), 99% (w / w), 99.5% (w / w), 99.8% (w / w), 99.9% (w / w), 100% (w / w), preferably 20% (w / w). Preferably, the lignans are selected from the above list. In a preferred embodiment, the determination of the polyphenol content is carried out using a method based on the Folin-Ciocalteu reagent (Singleton VL et al., 1999, Methods in enzimology, vol. 299: 152-78).

[0105] In another particular embodiment, any of the above extracts of one or more plant products of one or more plants of Polygonum cuspidatum, preferably the root extract, contains stilbene in an amount of 10% to 100% (w / w), 20% to 100% (w / w), 30% to 100% (w / w), 40% to 100% (w / w), 50% to 100% (w / w), 60% to 100% (w / w), 70% to 100% (w / w), 80% to 100% (w / w), 90% to 100% (w / w), 100% to 100% (w / w), 110% to 100% (w / w), 120% to 100% (w / w), 130% to 100% (w / w), 140% to 100% (w / w), 150% to 100% (w / w), 160% to 100% (w / w), 170% to 100% (w / w), 180% to 100% (w / w), 190% to 200% (w / w), 200% to 200% (w / w), 210% to 200% (w / w), 220% to 200% (w / w), 230% to 200% (w / w), 240% to 200% (w / w), 250% to 200% (w / w), 260% to 200% (w / w), 270% to 270% (w / w), 280% to 280% (w / w), 290% to 300% (w / w), 300% to 30 0% (w / w), 80%-100% (w / w), 85%-100% (w / w), 90%-99.9% (w / w), 90%-99.8% (w / w), 90%-99.5% (w / w), 90%-99.25% (w / w), 90%-99% (w / w), 92%-99% (w / w), 95%-99% (w / w), preferably 50%-100% (w / w) of stilbene. Preferably, said stilbene is selected from the list consisting of resveratrol, piceatannol, pinosylvin, pterostilbene, rhapontigenin and combinations thereof, more preferably resveratrol. In a particular embodiment, this is a combination of resveratrol with at least one of the other stilbenoids shown in the list above. In certain embodiments, the extract contains 50% (w / w), 60% (w / w), 65% (w / w), 70% (w / w), 75% (w / w), 80% (w / w), 85% (w / w), 90% (w / w), 91% (w / w), 92% (w / w), 93% (w / w), 94% (w / w), 95% (w / w), 96% (w / w), 100% (w / w), 102% (w / w), 104% (w / w), 105% (w / w), 106% (w / w), 107% (w / w), 108% (w / w), 109% (w / w), 110% (w / w), 111% (w / w), 112% (w / w), 113% (w / w), 114% (w / w), 115% (w / w), 116% (w / w), 117% (w / w), 118% (w / w), 119% (w / w), 120% (w / w), 121% (w / w), 122% (w / w), 123% (w / w), 124% (w / w), 125% (w / w), 126% (w / w), 127% (w / w), 128% (w / w), 129% (w / w), 130% (w / w), 131% (w / w), 132% (w / w), 133% (w / w), 134% (w / w), 135% (w / w), 136% (w / w), 137% (w / w), 138% (w / w), 139% (w / % (w / w), 97% (w / w), 98% (w / w), 99% (w / w), 99.5% (w / w), 99.75% (w / w), 99.8% (w / w), 99.85% (w / w), 99.9% (w / w), 99.95% (w / w), 99.99% (w / w), 100% (w / w), preferably 98% (w / w). Preferably, the stilbene is selected from the list consisting of resveratrol, piceatannol, pinosylvin, pterostilbene, rhapontigenin and combinations thereof, more preferably resveratrol. In a particular embodiment, it is a combination of resveratrol and at least one of the other stilbenoids from the list above.In a preferred embodiment, the determination of the polyphenol content is carried out using a method based on the Folin-Ciocalteu reagent (Singleton VL et al., 1999, Methods in Enzymology, vol. 299: 152-78).

[0106] In a preferred embodiment, the extract of Polygonum cuspidatum root contains 10% to 100% (w / w), 20% to 100% (w / w), 30% to 100% (w / w), 40% to 100% (w / w), 50% to 100% (w / w), 60% to 100% (w / w), 70% to 100% (w / w), 80% to 100% (w / w), %(w / w), 85%-100%(w / w), 90%-99.9%(w / w), 90%-99.8%(w / w), 90%-99.5%(w / w), 90%-99.25%(w / w), 90%-99%(w / w), 92%-99%(w / w), 95%-99%(w / w), preferably 50%-100%(w / w) of stilbene. Preferably, said stilbene is selected from the list consisting of resveratrol, piceatannol, pinosylvin, pterostilbene, rhapontigenin and combinations thereof, more preferably resveratrol. In a particular embodiment, this is a combination of resveratrol with at least one of the other stilbenoids shown in the list above. In certain embodiments, the extract contains 50% (w / w), 60% (w / w), 65% (w / w), 70% (w / w), 75% (w / w), 80% (w / w), 85% (w / w), 90% (w / w), 91% (w / w), 92% (w / w), 93% (w / w), 94% (w / w), 95% (w / w), 96% (w / w), 100% (w / w), 102% (w / w), 104% (w / w), 105% (w / w), 106% (w / w), 107% (w / w), 108% (w / w), 109% (w / w), 110% (w / w), 111% (w / w), 112% (w / w), 113% (w / w), 114% (w / w), 115% (w / w), 116% (w / w), 117% (w / w), 118% (w / w), 119% (w / w), 120% (w / w), 121% (w / w), 122% (w / w), 123% (w / w), 124% (w / w), 125% (w / w), 126% (w / w), 127% (w / w), 128% (w / w), 129% (w / w), 130% (w / w), 131% (w / w), 132% (w / w), 133% (w / w), 134% (w / w), 135% (w / w), 136% (w / w), 137% (w / w), 138% (w / w), 139% (w / % (w / w), 97% (w / w), 98% (w / w), 99% (w / w), 99.5% (w / w), 99.75% (w / w), 99.8% (w / w), 99.85% (w / w), 99.9% (w / w), 99.95% (w / w), 99.99% (w / w), 100% (w / w), preferably 98% (w / w). Preferably, the stilbene is selected from the list consisting of resveratrol, piceatannol, pinosylvin, pterostilbene, rhapontigenin and combinations thereof, more preferably resveratrol. In a particular embodiment, it is a combination of resveratrol and at least one of the other stilbenoids from the list above.In a preferred embodiment, the determination of the polyphenol content is carried out using a method based on the Folin-Ciocalteu reagent (Singleton VL et al., 1999, Methods in Enzymology, vol. 299: 152-78).

[0107] In a preferred embodiment, The extract of the pericarp of Prunus dulcis contains 5% to 60% (w / w), preferably 30% (w / w), of flavonoids, preferably selected from the group consisting of procyanidins, propelargonidins, prodelphinidins, catechins, epicatechins, quercetin, kaempferol, isorhamnetin, rutin, genistein, and combinations thereof. The Citrus Paradisi fruit extract or Citrus Paradisi fruit seed extract contains 10% to 80% w / w, preferably 45% (w / w), of flavonoids, preferably selected from the group consisting of naringin, naringenin, quercetin, catechin, rutin, genistein, and combinations thereof. The extract of the fruit seeds of Linum usitatissimum contains 5% to 50% (w / w), preferably 20% (w / w), of lignans, which are preferably selected from the group consisting of secoisolariciresinol diglucoside, secoisolariciresinol, matairesinoside, lariciresinol, pinoresinol, and sesamin, and combinations thereof. The Polygonum cuspidatum root extract contains 50% to 100% (w / w), preferably 98% (w / w), of stilbenoids, preferably selected from the group consisting of resveratrol, piceatannol, pinosylvin, pterostilbene, rhapontigenin, and combinations thereof.

[0108] In certain embodiments, the polyphenol other than hydroxytyrosol is at least 1.25 times, 1.5 times, 1.75 times, 2 times, 2.1 times, 2.2 times, 2.3 times, 2.4 times, 2.5 times, 2.6 times, 2.7 times, 2.8 times, 2.9 times, 3 times, 3.1 times, 3.2 times, 3.4 times, 3.5 times, 3.75 times, 4 times, 5 times, 6 times, 7 times, 8 times, 9 times, 10 times, 11 times, 12 times, 13 times, 14 times, 15 times, 16 times, 17 times, 18 times, 19 times, 20 times, 21 times, 22 times, 23 times, 24 times, 25 times, 26 times, 27 times, 28 times, 29 times, 30 times, 31 times, 32 times, 33 times, 34 times, 35 times, 36 times, 37 times, 38 times, 39 times, 40 times, 41 times, 42 times, 43 times, 44 times, 45 times, 46 times, 47 times, 48 ​​times, 49 times, 50 times, 51 times, 52 times, 53 times, 54 times, 55 times, 56 times, 57 times, 58 times, 59 times, 60 times, 61 times, 62 times, 63 times, 64 times, 65 times, 66 times, 67 times, 68 times, 69 times, 70 times, 71 times, 72 times, 73 times, 74 times, 75 times, 75 times, 76 times, 77 times, 78 times, 79 times, 8 , 4.25x, 4.5x, 4.75x, 5x, 5.5x, 6x, 6.5x, 7x, 7.5x, 8x, 8.5x, 9x, 10x, 11x, 12x, 51x, 18x, 20x, 21x, 22x, 23x, 24x, 24.2x, 24.3x, 24.5x, 25x, 26x, 27x, 28x, 29x, 30x, 32x, 35x, 40x, preferably 2.7x more is used.

[0109] In another particular embodiment, the w / w ratio of hydroxytyrosol to polyphenols other than hydroxytyrosol used in the first, second or third aspect is 1:0.5 to 1:100, 1:1 to 1:50, 1:2 to 1:40, 1:2 to 1:30, 1:2 to 1:28, 1:2 to 1:25, 1:2 to 1:24.5, 1:2 to 1:24.3, 1:2 to 1:24, 1:2 to 1:20, 1:2 to 1:17, 1:2 to 1:15, 1:2 to 1:10, 1:2 to 1:7, 1:2 to 1:5, 1:2 to 1:4, 1:2 to 1:3, preferably 1:1 to 1:50.

[0110] In another particular embodiment, the w / w ratio of hydroxytyrosol to polyphenols other than hydroxytyrosol used in the first, second or third aspect is at least 1:0.5, 1:1, 1:1.5, 1:2, 1:2.3, 1:2.5, 1:2.7, 1:2.8, 1:2.9, 1:3, 1:3.2, 1:3.5, 1:3.7, 1:4, 1:4.5, 1:5, 1:5.5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:12, 1:15, 1:17, 1:20, 1:21, 1:22, 1:23, 1:24, 1:24.1 , 1:24.2, 1:24.3, 1:24.4, 1:24.5, 1:24.6, 1:24.7, 1:24.8, 1:24.9, 1:25, 1:26, 1:27, 1:28, 1:29, 1:30, 1:32, 1:35, 1:37, 1:40, 1:45, 1:50, 1:55, 1:60, 1:65, 1:70, 1:80, 1:75, 1:80, 1:90, 1:100, 1:110, 1:120, 1:130, 1:140, 1:150, 1:200, preferably 1:2.7, more preferably 1:24.3, and even more preferably 1:28.

[0111] In certain embodiments, the w / w ratio of plant extract containing hydroxytyrosol to plant extract containing at least one polyphenol other than hydroxytyrosol used in the first, second or third aspect is 1:0.5 to 1:100, 1:1 to 1:75, 1:2 to 1:50, 1:3 to 1:40, 1:4 to 1:30, 1:5 to 1:25, 1:7 to 1:20, 1:10 to 1:15, preferably 1:2 to 1:50. In another particular embodiment, the w / w ratio of plant extract containing hydroxytyrosol to plant extract containing polyphenols other than hydroxytyrosol is at least 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:17, 1:20, 1:22, 1:25, 1:27, 1:30, 1:35, 1:40, 1:45, 1:50, 1:55, 1:60, 1:70, 1:80; 1:90, 1:100.

[0112] 2. Compositions and kits of parts of the present invention In a fourth aspect, the present invention relates to a composition or kit of parts comprising hydroxytyrosol and at least one polyphenol that is not hydroxytyrosol, at least one polyphenol that does not naturally occur in the O. europaea tree, or at least one polyphenol that is other than hydroxytyrosol and that does not naturally occur in the O. europaea tree.

[0113] In certain embodiments, hydroxytyrosol and at least one polyphenol that is not hydroxytyrosol, at least one polyphenol that does not naturally occur in the O. europaea tree, or at least one polyphenol other than hydroxytyrosol and that does not naturally occur in the O. europaea tree are referred to as compounds of the compositions of the present invention.

[0114] As used in the present invention, the term "composition" refers to a combination of compounds or components. The components of the composition can be provided separately or in one dosage form. Thus, when the composition is administered to a subject, the compounds contained in the composition can be mixed in the dosage form or can be mixed before administration, and whether they are provided separately or provided and administered separately, they are mixed once ingested by the subject, i.e., mixed in the subject's body. Furthermore, some of the components of the composition can be administered together and others can be administered separately, but they are all mixed once ingested by the subject, i.e., mixed in the subject's body.

[0115] The composition according to the present invention can be provided in various forms. Examples include, but are not limited to, tablets, coated tablets, pills, aqueous or oily suspensions, liquids, dispersible powders or granules, emulsions, hard or soft capsules, syrups or elixirs, pastes, gels, etc. The compositions can be prepared by any known method, and such compositions can contain, in addition to the compounds described above in the compositions of the present invention, one or more agents that provide a pharmacopoeial and pleasant composition, the agents being selected from the group consisting of sweeteners, flavoring agents, coloring agents, and preservatives. Regardless of the form in which the composition is provided, the active ingredient(s) can be contained in a mixture with non-toxic pharmacopoeiacally acceptable excipients suitable for the manufacture of the form of delivery. Such excipients can be, for example, inert diluents such as calcium carbonate, sodium carbonate, lactose, calcium phosphate, or sodium phosphate; granulating and disintegrating agents such as corn starch or alginic acid; binders such as starch, gelatin, or gum acacia; and lubricants such as magnesium stearate, stearic acid, or talc. The various forms in which the composition is delivered can be uncoated or coated by known techniques to delay disintegration and absorption in the digestive system, thereby providing a sustained action over a long period of time. For example, a time-delay material such as glyceryl monostearate or glyceryl distearate can be used. Such forms can also be coated for controlled administration. For example, a "slow-release" dosage form releases a product or substance at different times from immediately after administration. Examples of slow-release systems include reactivation tablets and capsules, and enteric-coated tablets, where a scheduled release is achieved through a barrier coating.

[0116] Compositions according to the invention may also be formulated for oral use as hard gelatin capsules in which the active ingredient(s) are mixed with an inert solid diluent, such as calcium carbonate, calcium phosphate, or kaolin, or as soft gelatin capsules in which the active ingredient(s) are mixed with water and an oil medium, such as peanut oil, liquid paraffin, or olive oil.

[0117] The composition according to the invention can be formulated as an aqueous suspension, in which the active ingredient(s) are mixed with excipients suitable for the manufacture of aqueous suspensions. Such excipients include suspending agents, such as sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, gum tragacanth, and gum acacia. Dispersing or wetting agents can be natural phosphatides, such as lecithin, or condensation products of alkylene oxides with fatty acids, such as polyoxyethylene stearates, or condensation products of ethylene oxide with long-chain aliphatic alcohols, such as heptadecaethylenoxyketanol, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitols, such as polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, such as polyethylene sorbitan monooleate. The aqueous suspensions may also contain one or more colouring agents, one or more flavouring agents, and one or more sweetening agents, such as sucrose or saccharin.

[0118] The compositions according to the invention can be formulated as oily suspensions by suspending the active ingredient in a vegetable oil, such as peanut oil, olive oil, sesame seed oil, or coconut oil, or in a mineral oil, such as liquid paraffin. Oily suspensions can contain thickening agents, such as beeswax, hard paraffin, or cetyl alcohol. Sweeteners and flavoring agents, such as those listed above, can be added to provide an orally acceptable composition. These compositions can be preserved by adding an antioxidant, such as ascorbic acid.

[0119] The composition according to the present invention can be formulated into dispersible powder and granule form suitable for adding water to make aqueous suspension composition.The active ingredient in such powder and granule is provided as a mixture with dispersing or wetting agent, suspending agent, and one or more preservatives.The dispersing or wetting agent and suspending agent are exemplified by those already mentioned above.Additional excipients, such as sweeteners, flavoring agents, and coloring agents, can also be present.

[0120] The composition according to the invention can be in the form of an oil-in-water emulsion.The oil phase can be a vegetable oil, for example olive oil or peanut oil, or a mineral oil, for example liquid paraffin, or a mixture thereof.Suitable emulsifiers can be natural gums, for example gum acacia or gum tragacanth, natural phosphatides, for example soybean lecithin, and esters or partial esters derived from fatty acids and hexitol anhydrides, for example sorbitan monooleate, and condensation products of partial esters with ethylene oxide, for example polyoxyethylene sorbitan monooleate, etc.The emulsion can also contain sweeteners and flavoring agents.

[0121] The compositions according to the invention can be formulated as syrups and elixirs. Syrups and elixirs can be formulated with sweeteners, such as glycerol, propylene glycol, sorbitol, or sucrose. Such formulations can also contain demulcents, preservatives, and flavoring and coloring agents. Demulcents are protective agents primarily used to relieve irritation, especially to mucous membranes or degraded tissue. Several chemicals possess demulcent properties. Such substances include alginic acid compounds, mucilages, gums, dextrins, starches, certain sugars, and polymerized polyhydric glycols. Others include gum acacia, agar, benzoin, carbomer, gelatin, glycerin, hydroxyethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methylcellulose, propylene glycol, sodium alginate, tragacanth, hydrogels, and the like.

[0122] Suitable formulations for oral administration include tablets and coated tablets containing the compounds of the composition of the present invention in a flavored base such as sucrose, acacia, or tragacanth, and pills containing the compounds in an inert base such as gelatin and glycerin or sucrose and acacia.

[0123] Those skilled in the art will be able to appropriately formulate liquid formulations of the compositions of the present invention, and their solid analogs, which will contain appropriate amounts of each of the compounds of the compositions of the present invention, depending on the additive or support selected.

[0124] As used in the present invention, the expression "kit of parts" refers to a product that includes various ingredients, components, or compounds that are physically separated, preferably by packaging each ingredient, component, or compound in the kit separately, in a manner that allows for transport and storage. It is understood that in a "kit of parts" according to the present invention, the individual active ingredients, components, or compounds are therapeutic agents, provided that the use of these compounds, whether simultaneously, separately, or sequentially, together, as described herein, provides a new and unexpected therapeutic effect that cannot be achieved by each compound independently relative to each other. Indeed, as will be demonstrated by the results below, the claimed combination of active ingredients was not a mere collection of known agents. Instead, it is a novel combination with beneficial and surprising properties, the combined effect of which is much more significant than the simple sum of the effects observed when these active ingredients are used separately. A kit of parts typically contains its components in a suitable container. Suitable materials for packaging the components of the kit include glass, plastic (polyethylene, polypropylene, polycarbonate, etc.), bottles, vials, paper, pouches, etc. For example, each container can be a vial, bottle, screw-on bottle, jar, closure sleeve, sachet or pouch, tube or blister, or any other suitable shape, provided that the container is configured to prevent incomplete mixing of the components. Each of the different components can be provided separately, or portions of the different components can be provided together (i.e., in the same container). Furthermore, the kits of the invention can include instructions for the simultaneous, sequential, or separate use of the different components included in the kit. The instructions can be in the form of printed matter or in the form of an electronic medium on which the instructions can be recorded such that the subject can read them, such as an electronic recording medium (magnetic disk, tape, etc.), optical medium (CD-ROM, DVD), etc. The medium can additionally or alternatively include an internet address providing the instructions.

[0125] It will be appreciated that the compositions or kits of parts of the present invention may comprise, consist essentially of, or consist of the above-mentioned components, compounds, or ingredients.

[0126] In this specification, the term "comprising" or "comprises" is used to indicate that the composition described must contain the recited component(s), but the composition may optionally contain additional components. The term "essentially consisting of" or "essentially consists of" is used to indicate that the composition described must contain the recited component(s), but the composition may also contain other components in small amounts (e.g., up to 5% by weight, or up to 1% by weight, or 0.1% by weight), but without additional components that affect the essential nature of the extract or composition. The term "consisting of" or "consists of" is used to indicate that the composition described must contain only the recited component(s).

[0127] In certain embodiments, the composition or kit of parts according to the fourth aspect of the invention comprises, in addition to hydroxytyrosol and at least one polyphenol other than hydroxytyrosol, additional components. In certain embodiments, this comprises at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, at least 15, at least 20, at least 25, at least 30, at least 40, at least 50, at least 60, at least 70, at least 80, at least 90, at least 100 components in addition to hydroxytyrosol and at least one polyphenol other than hydroxytyrosol. In another specific embodiment, hydroxytyrosol and at least one polyphenol other than hydroxytyrosol constitute at least 0.25%, at least 0.5%, at least 1%, at least 2%, at least 3%, at least 4%, at least 5%, at least 6%, at least 7%, at least 8%, at least 9%, at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, at least 50%, at least 55%, at least 60%, at least 65%, at least 66%, at least 67%, at least 70%, at least 75%, at least 80%, at least 85%, or at least 90%, at least 100% of the total amount of the kit.

[0128] The term "polyphenol" and the expression "a polyphenol that is not hydroxytyrosol, in this case at least one polyphenol other than hydroxytyrosol and / or that is not naturally occurring in the O. europaea tree" are defined in the first, second and third aspects of the invention. The definitions and the specific embodiments describing the terms and expressions apply to this aspect of the invention.

[0129] In certain embodiments, the hydroxytyrosol is part of a plant extract.

[0130] Thus, in certain embodiments, as will be appreciated by those skilled in the art, the "hydroxytyrosol" compound or component of the compositions and kits of parts of the fourth aspect of the invention consists of a plant extract that contains hydroxytyrosol.

[0131] The expressions "plant extract" and "part of a plant extract" are defined in the first, second and third aspects of the invention. Thus, as will be appreciated by the skilled artisan, in certain embodiments, hydroxytyrosol is provided in the composition of the fourth aspect of the invention as a plant extract.

[0132] In certain embodiments, the hydroxytyrosol-containing plant extract of this aspect of the invention is similar to the hydroxytyrosol-containing extract described in any of the embodiments and definitions of the first, second and third aspects of the invention.

[0133] Thus, in certain embodiments, the plant extract comprising hydroxytyrosol is obtained from any plant product of the O. europaea tree, which plant product is any of those set out in the definition of "plant product" set out in the first, second or third aspect of the present invention.

[0134] In a preferred embodiment of the fourth aspect of the invention, the plant extract of which hydroxytyrosol is a part is an extract of the leaves or fruits, preferably the leaves, of the O. europaea tree.

[0135] In a particular embodiment, the extract is any extract mentioned in the first, second and third aspects of the invention of one or more plant products of one or more trees of O. europaea, preferably an extract of leaves or fruits, more preferably an extract of leaves.

[0136] In certain embodiments, the hydroxytyrosol content in the hydroxytyrosol-containing extract of this aspect is any of those set out for the hydroxytyrosol-containing extract of the aspect in the first, second and third aspects of the invention.

[0137] In a preferred embodiment of the fourth aspect of the present invention, the hydroxytyrosol-containing plant extract contains 10% to 90% hydroxytyrosol, preferably 10% (w / w).

[0138] In another particular embodiment, the at least one non-hydroxytyrosol polyphenol of the fourth aspect of the invention is at least one non-hydroxytyrosol polyphenol as defined and described in the first, second and third aspects of the invention. Thus, the non-hydroxytyrosol polyphenol of the fourth aspect of the invention is any of the polyphenols defined as belonging to the group of non-hydroxytyrosol polyphenols as set forth in the first, second and third aspects of the invention.

[0139] In a preferred embodiment, the at least one polyphenol other than hydroxytyrosol is selected from the group of polyphenols consisting of flavonoids, lignans, and stilbenes.

[0140] In another particular embodiment, the at least one polyphenol other than hydroxytyrosol from the flavonoid group is selected from the group consisting of procyanidin, propelargonidin, prodelphinidin, catechin, epicatechin, quercetin, kaempferol, isorhamnetin, naringin, naringenin, genistein, and rutin.

[0141] In a preferred embodiment, the procyanidin is a B-type procyanidin, preferably selected from the group consisting of B1-type procyanidin, B2-type procyanidin, B3-type procyanidin, B4-type procyanidin, B5-type procyanidin, B6-type procyanidin, and B7-type procyanidin. In another particular embodiment, it is a C1-type procyanidin. In a particular embodiment, it is an A-type procyanidin, preferably selected from the group consisting of procyanidin A1, procyanidin A2.

[0142] In another specific embodiment, the propelargonidin is type A, more preferably type B propelargonidin.

[0143] In certain embodiments, the prodelphinidin is type A.

[0144] In another particular embodiment, the at least one polyphenol other than hydroxytyrosol from the lignan group is selected from the group consisting of secoisolariciresinol diglucoside, secoisolariciresinol, matairesinoside, lariciresinol, pinoresinol, and sesamin.

[0145] In another particular embodiment, the at least one polyphenol other than hydroxytyrosol from the stilbene group is selected from the group consisting of resveratrol, piceatannol, pinosylvin, pterostilbene and rhapontigenin, preferably resveratrol.

[0146] In another particular embodiment of the composition or kit of parts of the invention, the at least one polyphenol other than hydroxytyrosol is part of a plant extract.

[0147] In another particular embodiment, as will be appreciated by those skilled in the art, the "polyphenol other than hydroxytyrosol" component or compound of the composition or kit of parts of the fourth aspect of the invention consists of a plant extract.

[0148] In another particular embodiment, the plant extract of which at least one polyphenol other than hydroxytyrosol is a part is not an extract of a plant product of the O. europaea tree, and preferably it is not an extract of the leaves or fruits of the O. europaea tree.

[0149] In another particular embodiment, the plant extract of which at least one polyphenol other than hydroxytyrosol is a part is as defined and described in the first, second and third aspects of the invention.

[0150] Thus, in a preferred embodiment, the plant extract of which at least one polyphenol other than hydroxytyrosol is a part is an extract selected from the group consisting of: Prunus dulcis peel extract Citrus paradisi fruit extract or Citrus paradisi fruit seed extract Linum usitatissimum fruit seed extract Polygonum cuspidatum root extract

[0151] In certain embodiments, the at least one polyphenol other than hydroxytyrosol present in each extract comprising other polyphenols of this aspect of the invention is as set out for that extract in the first, second or third aspect of the invention.

[0152] In a fourth preferred embodiment of the present invention, The at least one polyphenol other than hydroxytyrosol that is part of the extract of the pericarp of Prunus dulcis is a flavonoid, preferably selected from the group consisting of procyanidin, propelargonidin, prodelphinidin, catechin, epicatechin, quercetin, kaempferol, isorhamnetin, rutin, and genistein. At least one polyphenol other than hydroxytyrosol that is part of the Citrus Paradisi fruit extract or Citrus Paradisi fruit seed extract is a flavonoid, preferably selected from the group consisting of naringin, naringenin, quercetin, catechin, rutin, and genistein. The at least one polyphenol other than hydroxytyrosol which is part of the extract of the seeds of the fruit of Linum usitatissimum is a lignan, preferably selected from the group consisting of secoisolariciresinol diglucoside, secoisolariciresinol, matairesinoside, lariciresinol, pinoresinol, and sesamin. The at least one polyphenol other than hydroxytyrosol that is part of the Polygonum cuspidatum root extract is a stilbene, preferably selected from the group consisting of resveratrol, piceatannol, pinosylvin, pterostilbene, and rhapontigenin, more preferably resveratrol.

[0153] In a particular embodiment, the content and type of flavonoids contained in any of the extracts referred to in the present invention, preferably an extract of the pericarp, of one or more plant products of one or more trees of Prunus dulcis, are as indicated for said extracts in the first, second and third aspects of the present invention.

[0154] In another particular embodiment, the content and type of flavonoids contained in any extract mentioned in the present invention of one or more plant products of one or more Citrus paradisi trees, preferably an extract of the fruits or seeds of the fruits, are as indicated for said extracts in the first, second and third aspects of the present invention.

[0155] In another particular embodiment, the content and type of lignans contained in any extract mentioned in the present invention of one or more plant products of one or more plants of Linum usitatissimum, preferably an extract of the fruit seeds, are as indicated for said extracts in the first, second and third aspects of the present invention.

[0156] In another particular embodiment, the content and type of stilbenes contained in any of the extracts referred to in the present invention, preferably the root extract, of one or more plant products of Polygonum cuspidatum are as indicated for said extracts in the first, second and third aspects of the present invention.

[0157] Thus, in a preferred embodiment, the extract of the pericarp of Prunus dulcis contains 5% to 60% (w / w) of flavonoids, preferably 30% (w / w), the flavonoids being preferably selected from the group consisting of procyanidins, propelargonidins, prodelphinidins, catechins, epicatechins, quercetin, kaempferol, isorhamnetin, rutin, genistein, and combinations thereof. The Citrus Paradisi fruit extract or Citrus Paradisi fruit seed extract contains 10% to 80% w / w, preferably 45% (w / w), of flavonoids, preferably selected from the group consisting of naringin, naringenin, quercetin, catechin, rutin, genistein, and combinations thereof. The extract of the fruit seeds of Linum usitatissimum contains 5% to 50% (w / w), preferably 20% (w / w), of lignans, which are preferably selected from the group consisting of secoisolariciresinol diglucoside, secoisolariciresinol, matairesinoside, lariciresinol, pinoresinol, and sesamin, and combinations thereof. The Polygonum cuspidatum root extract contains 50% to 100% (w / w), preferably 98% (w / w), of stilbenoids, preferably selected from the group consisting of resveratrol, piceatannol, pinosylvin, pterostilbene, rhapontigenin, and combinations thereof.

[0158] In certain embodiments, in the composition or kit of parts of the fourth aspect of the invention, the polyphenol other than hydroxytyrosol is present in an amount of at least 1.25x, 1.5x, 1.75x, 2x, 2.1x, 2.2x, 2.3x, 2.4x, 2.5x, 2.6x, 2.7x, 2.8x, 2.9x, 3x, 3.1x, 3.2x, 3.4x, 3.5x, 3.75x, 4x, 4.25x, 4.5x, 4.75x, 5x, 5.5x, 6x, 7x, 8x, 9x, 10x, 11x, 12x, 13x, 14x, 15x, 16x, 17x, 18x, 19x, 20x, 21x, 22x, 23x, 24x, 25x, 26x, 27x, 28x, 29x, 30x, 31x, 32x, 33x, 34x, 35x, 36x, 37x, 38x, 39x, 40x, 41x, 42x, 43x, 44x, 45x, 46x, 47x, 48x, 49x, 50x, 51x, 52x, 53x, 54x, 55x, 56x, 57x, 58x, 59x, 60x, 61x, 62x, 63x, 64x, 65x, 66x, 67x, 68x, 69x, 70x, 71x, 6x, 6.5x, 7x, 7.5x, 8x, 8.5x, 9x, 10x, 11x, 12x, 51x, 18x, 20x, 21x, 22x, 23x, 24x, 24.2x, 24.3x, 24.4x, 24.5x, 25x, 26x, 27x, 28x, 29x, 30x, 32x, 35x, 40x, 54x, 50x, 55x, 60x, 65x, 70x, 80x, 85x, 90x, 95x, 100x, preferably 2.7x more, more preferably 24.3x more by weight, and even more preferably at least 28x more by weight.

[0159] In another particular embodiment of the fourth aspect of the invention, the w / w ratio of hydroxytyrosol:polyphenols other than hydroxytyrosol in the composition or kit of parts is between 1:0.5 and 1:100, between 1:1 and 1:50, between 1:2 and 1:40, between 1:2 and 1:30, between 1:2 and 1:28, between 1:2 and 1:25, between 1:2 and 1:24.5, between 1:2 and 1:24.3, between 1:2 and 1:24, between 1:2 and 1:20, between 1:2 and 1:17, between 1:2 and 1:15, between 1:2 and 1:10, between 1:2 and 1:7, between 1:2 and 1:5, between 1:2 and 1:4, between 1:2 and 1:3, preferably between 1:1 and 1:30. In another particular embodiment, the w / w ratio of hydroxytyrosol:polyphenols other than hydroxytyrosol in the composition or kit of parts is 1:0.5, 1:1, 1:1.5, 1:2, 1:2.3, 1:2.5, 1:2.7, 1:2.8, 1:2.9, 1:3, 1:3.2, 1:3.5, 1:3.7, 1:4, 1:4.5, 1:5, 1:5.5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:12, 1:15, 1:17, 1:20, 1:21, 1:22, 1:23, 1:24, 1:24.1, 1:24.2, 1:24.3, 1:24.4, 1:24.5, 1:24.6, 1:24.7, 1:24.8, 1:24.9, 1:25, 1:26, 1:27, 1:28, 1:29, 1:30, 1:32, 1:35, 1:37, 1:40, 1:45, 1:50, 1:55, 1:60, 1:65, 1:70, 1:80, 1:75, 1:80, 1:90, 1:100, 1:110, 1:120, 1:130, 1:140, 1:150, 1:200, preferably 1:2.7, more preferably 1:24.3, even more preferably 1:28.

[0160] In a particular embodiment, in the composition or kit of parts of the fourth aspect of the invention, the w / w ratio of hydroxytyrosol:polyphenols other than hydroxytyrosol is between 1:1 and 1:50, preferably 1:24.3, more preferably 1:28.

[0161] In certain embodiments, in the composition or kit of parts of the fourth aspect of the invention, the w / w ratio of plant extract containing hydroxytyrosol to plant extract containing polyphenols other than hydroxytyrosol is between 1:0.5 and 1:100, 1:1: to 1:75, 1:2 to 1:50, 1:3 to 1:40, 1:4 to 1:30, 1:5 to 1:25, 1:7 to 1:20, 1:10 to 1:15, preferably between 1:5 and 1:25. In another particular embodiment, the w / w ratio of plant extract containing hydroxytyrosol to plant extract containing polyphenols other than hydroxytyrosol is at least 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:11, 1:12, 1:13, 1:14, 1:15, 1:17, 1:20, 1:22, 1:25, 1:27, 1:30, 1:35, 1:40, 1:45, 1:50, 1:55, 1:60, 1:70, 1:80; 1:90, 1:100, preferably at least 1:10.

[0162] In another particular embodiment, the hydroxytyrosol content in the composition of the fourth aspect of the invention is at least 0.02% to 20% (w / w), at least 0.025% to 15% (w / w), at least 0.5% to 10% (w / w), at least 0.5% to 7.5% (w / w), at least 0.6% to 5% (w / w), at least 0.7% to 2.5% (w / w), at least 0.8% to 2% (w / w), at least 0.9% to 1.5% (w / w), more preferably at least 0.05% to 10% (w / w), based on the total composition. In another particular embodiment, the almond skin polyphenol composition in the composition of the fourth aspect of the invention is at least 5% (w / w), at least 7% (w / w), at least 10% (w / w), at least 15% (w / w), at least 20% (w / w), at least 30% (w / w), preferably at least 10% (w / w), based on the total composition. In another particular embodiment, the almond skin polyphenol composition in the composition of the fourth aspect of the invention is at least 5% (w / w), at least 7% (w / w), at least 10% (w / w), at least 12% (w / w), at least 15% (w / w), at least 17% (w / w), at least 20% (w / w), at least 21% (w / w), at least 22% (w / w), at least 23% (w / w), at least 24% (w / w), based on the total composition. / w), at least 25% (w / w), at least 26% (w / w), at least 27% (w / w), at least 28% (w / w), at least 29% (w / w), at least 30% (w / w), at least 32% (w / w), at least 35% (w / w), at least 40% (w / w), at least 45% (w / w), at least 50% (w / w), at least 55% (w / w), at least 60% (w / w), preferably at least 26% (w / w).

[0163] In a preferred embodiment, the hydroxytyrosol content in the composition according to the fourth aspect of the invention is at least 0.05% to 10% w / w, preferably at least 0.9% w / w, based on the total composition, and the at least one polyphenol other than hydroxytyrosol in the composition is an almond skin polyphenol composition, the content of which is at least about 10% to 50% w / w, preferably at least 26% w / w, based on the total composition.

[0164] In certain embodiments, the definitions and specific embodiments of the first, second and third aspects of the invention apply to the fourth aspect of the invention.

[0165] 3. Methods of the Invention In a fifth aspect, the present invention relates to a method of obtaining a composition according to the fourth aspect of the invention, comprising contacting a composition comprising hydroxytyrosol with a composition comprising at least one polyphenol which is not hydroxytyrosol and which does not naturally occur in the O. europaea tree.

[0166] The term "composition" is defined in the fourth aspect of the invention. The terms "hydroxytyrosol" and "non-hydroxytyrosol polyphenols" are defined in the first, second and third aspects of the invention.

[0167] In certain embodiments, the polyphenol other than hydroxytyrosol is as defined in any of the definitions and in certain embodiments of the first, second, third and fourth aspects of the invention.

[0168] As used herein, the term "contacting" refers to a mixture or combination of a composition comprising hydroxytyrosol and a composition comprising a polyphenol other than hydroxytyrosol, such that the components of one composition are surrounded by the components of the other composition. As a result, a composition is obtained that comprises a composition comprising hydroxytyrosol and a composition comprising a polyphenol other than hydroxytyrosol. The product obtained by said contacting is the composition of the first, second, third or fourth aspect of the present invention.

[0169] The contacting can be performed in vitro, as described below, or after the subject ingests a composition containing hydroxytyrosol and a composition containing polyphenols other than hydroxytyrosol, either simultaneously or separately, in which case contact between the two compositions occurs once they enter the subject's body.

[0170] A mixture or combination of a composition containing hydroxytyrosol and a composition containing a polyphenol other than hydroxytyrosol can be carried out by placing both compositions in the same container and moving the contents of the container so that the components of one composition are between the components of the other composition with a certain uniformity. In a specific embodiment, the mixture contains at least one non-toxic pharma- ceutically acceptable excipient suitable for producing the composition of the first, second, third or fourth aspect of the present invention. The excipient can be any of the inert diluents, excipients suitable for producing aqueous suspensions, vegetable oils, thickeners, antioxidants, dispersants or wetting agents, suspending agents, emulsifiers, sweeteners or flavoring agents shown in the fourth aspect as being capable of being contained in the composition of the aspect.

[0171] A person skilled in the art will be able to appropriately formulate a mixture containing an appropriate amount of a composition comprising hydroxytyrosol and an appropriate amount of a composition comprising a polyphenol other than hydroxytyrosol depending on the excipient or additive selected.

[0172] In certain embodiments, the composition comprising hydroxytyrosol is a plant extract. In another particular embodiment, the composition comprising at least one polyphenol other than hydroxytyrosol is a plant extract.

[0173] In a preferred embodiment, the composition comprising hydroxytyrosol is a plant extract and / or the composition comprising at least one polyphenol that is not hydroxytyrosol is a plant extract.

[0174] The hydroxytyrosol-containing plant extract is the same as the hydroxytyrosol-containing plant extract defined in any of the above aspects of the invention.

[0175] In a preferred embodiment, the plant extract containing hydroxytyrosol is an extract of the leaves or fruits of the O. europaea tree, preferably a leaf extract.

[0176] In certain embodiments, the hydroxytyrosol content in the extract comprising hydroxytyrosol is any of the hydroxytyrosol contents in said extract as set out in any of the above aspects of the invention.

[0177] In another particular embodiment, the hydroxytyrosol-containing plant extract contains 1% to 90% w / w hydroxytyrosol, preferably 10% (w / w).

[0178] With regard to plant extracts comprising at least one polyphenol other than hydroxytyrosol, this is similar to any extract comprising at least one polyphenol other than hydroxytyrosol as defined in any of the above aspects of the invention.

[0179] In a preferred embodiment, the plant extract comprising at least one polyphenol that is not hydroxytyrosol is selected from the group consisting of: Prunus dulcis peel extract, Citrus paradisi fruit extract or Citrus paradisi fruit seed extract Extract of the seeds of the fruit of Linum usitatissimum, and Polygonum cuspidatum root extract.

[0180] In a particular embodiment, the content and type of flavonoids contained in any of the extracts referred to in the present invention, preferably an extract of the pericarp, of one or more plant products of one or more trees of Prunus dulcis, are as indicated for said extracts in the first, second and third aspects of the present invention.

[0181] In another particular embodiment, the content and type of flavonoids contained in any extract mentioned in the present invention of one or more plant products of one or more Citrus paradisi trees, preferably an extract of the fruits or seeds of the fruits, are as indicated for said extracts in the first, second and third aspects of the present invention.

[0182] In another particular embodiment, the content and type of lignans contained in any extract mentioned in the present invention of one or more plant products of one or more plants of Linum usitatissimum, preferably an extract of the fruit seeds, are as indicated for said extracts in the first, second and third aspects of the present invention.

[0183] In another particular embodiment, the content and type of stilbenes contained in any of the extracts referred to in the present invention, preferably the root extract, of one or more plant products of Polygonum cuspidatum are as indicated for said extracts in the first, second and third aspects of the present invention.

[0184] In certain embodiments, The extract of the pericarp of Prunus dulcis contains 5% to 60% w / w of flavonoids, preferably 30% w / w, the flavonoids being preferably selected from the group consisting of procyanidins, propelargonidins, prodelphinidins, catechins, epicatechins, quercetin, kaempferol, isorhamnetin, rutin, genistein, and combinations thereof. The Citrus Paradisi fruit extract or Citrus Paradisi fruit seed extract contains 10% to 80% w / w, preferably 45% (w / w) flavonoids, preferably selected from the group consisting of naringin, naringenin, quercetin, catechin, rutin, genistein, and combinations thereof. The extract of the fruit seeds of Linum usitatissimum contains 5% to 50% w / w, preferably 20% (w / w), of lignans, which are preferably selected from the group consisting of secoisolariciresinol diglucoside, secoisolariciresinol, matairesinoside, lariciresinol, pinoresinol, and sesamin. The Polygonum cuspidatum root extract contains 50%-100% w / w, preferably 98% (w / w) of stilbene, preferably selected from the group consisting of resveratrol, piceatannol, pinosylvin, pterostilbene, rhapontigenin, and combinations thereof, preferably resveratrol, and more preferably resveratrol in combination with at least one of the stilbenes listed above.

[0185] In certain embodiments, the definitions and particular embodiments of the first, second, third and fourth aspects of the invention apply to the fifth aspect of the invention.

[0186] 4. Pharmaceutical Composition of the Present Invention In a sixth aspect, the present invention relates to a pharmaceutical composition comprising the composition of the fourth aspect of the invention or the composition obtainable by the method of the fifth aspect of the invention, and a pharma- ceutically acceptable excipient.

[0187] As used herein, the expression "pharmaceutical composition" refers to any physiologically tolerable composition which, when administered to humans or animals, typically does not produce allergic reactions or similar untoward reactions, such as gastric upset, dizziness, etc.

[0188] The composition comprises at least one pharma- ceutical active ingredient and one or more pharma- ceutical acceptable adjuvants. The terms "pharma- ceutical acceptable adjuvants", "pharma- ceutical acceptable excipients", "pharma- ceutical acceptable diluents" or "pharma- ceutical acceptable vehicles" are used interchangeably herein and refer to non-toxic fillers, diluents, auxiliary agents, or encapsulating materials of any conventional solid, semi-solid, or liquid formulation. Pharmaceutically acceptable adjuvants are essentially non-toxic to the recipient at the dosages and concentrations used and are compatible with other ingredients of the formulation. Suitable adjuvants include, but are not limited to, water, glucose, glycerol, saline, ethanol, and combinations thereof. Adjuvants may contain additional agents, such as wetting or emulsifying agents, pH adjusting agents, or adjuvants that improve the effectiveness of the formulation. The adjuvant can be selected from the group consisting of sterile liquids such as water and oils, including those of petroleum, animal, vegetable, or synthetic origin, e.g., peanut oil, soybean oil, petrolatum oil, sesame seed oil, and the like. Water or aqueous saline solutions and aqueous dextrose and glycerol solutions, especially those for injection, are suitably used as vehicles. Suitable pharmaceutical vehicles are described in "Remington's Pharmaceutical Sciences" by EW Martin, 21 st Edition, 2005. The use of conventional adjuvants in therapeutic or pharmaceutical compositions is contemplated, except where such adjuvants are incompatible with the active ingredient. Pharmaceutically acceptable excipients also include any of the inert diluents shown in the fourth embodiment as compositions of the embodiment may contain, excipients suitable for the manufacture of aqueous suspensions, vegetable oils, thickeners, antioxidants, dispersing or wetting agents, suspending agents, emulsifying agents, sweeteners, or flavoring agents.

[0189] The pharmaceutical composition comprises a therapeutically effective amount of the plant extract of any of the aspects of the invention. As used herein, the term "effective amount" is synonymous with "therapeutically effective amount", "effective dose" or "therapeutically effective dose", and as used herein, the term refers to the minimum dose of the plant extract of any of the aspects of the invention required to achieve the desired therapeutic effect, including a dose sufficient to reduce at least one cardiovascular disease symptom, preferably atherosclerosis. The efficacy in treating the diseases or conditions described herein can be determined by observing a decrease in serum oxLDL levels and oxLDL / LDL ratio. In certain embodiments, the efficacy in treating the diseases or conditions described herein is as described in any of the specific embodiments given for the expression "increasing the anti-atherosclerotic effect of hydroxytyrosol" in the first, second or third aspects of the invention. In that case, the relevant reference value can be the one indicated in the embodiment, or the value of the relevant parameter in a serum sample from a subject not receiving any kind of treatment, or the value of the relevant parameter in a serum sample from the same subject to which the pharmaceutical composition is administered, obtained at least 2 weeks or at least 4 weeks prior to said administration. Thus, the method of measuring the efficacy in the treatment of a disease or condition described herein is also described in the embodiment of the first, second or third aspect of the invention.

[0190] A person skilled in the art can identify therapeutically effective amounts of the compositions of the present invention when administered in unit doses. As used herein, "unit dose" refers to the amount of the compositions or kits of parts of the present invention required to produce a 50 percent maximum response (i.e., ED50). Unit doses can be estimated by extrapolation of dose-response curves derived from in vitro or animal model test systems. The amount of compound in the compositions of the present invention that is effective in treating a particular disorder or condition will depend on the nature of the disorder or condition and can be identified by standard clinical techniques (see, for example, Goodman and Gilman's The Pharmacological Basis of Therapeutics, Joel G. Harman, Lee E. Limbird, Eds., McGraw Hill, Nueva York, 2001; The Physician's Desk Reference, Medical Economics Company, Inc., Oradell, NJ, 1995; and Drug Facts and Comparisons, Facts and Comparisons, Inc., St. Luis, Mo., 1993). The precise dose to be employed in the formulation will also depend on the route of administration, and the disorder or its seriousness, and must be decided according to the evaluation of the health care professional and each patient's circumstances. Multiple administration patterns will be apparent to those skilled in the art.

[0191] Furthermore, when repeated administration of any of the compounds of the above aspects of the invention (i.e., hydroxytyrosol, plant extracts containing hydroxytyrosol, polyphenols other than hydroxytyrosol, and / or extracts containing polyphenols other than hydroxytyrosol) is used, the effective amount of the compound will further depend on several factors, including, but not limited to, frequency of administration, half-life of the compound, or any combination thereof.

[0192] The dosage range of the composition of the present invention is large enough to produce the desired therapeutic effect. Preferably, the composition of the present invention is administered once or twice a day on a regular basis. The typical dose administered to a human is about 20 mg to about 5 g of the composition, preferably 200 mg to 1 g of the composition.

[0193] The compositions provided herein can be administered to a subject by a number of suitable methods known in the art.Suitable methods include: (1) intramuscular, intradermal, intraepidermal, or subcutaneous administration, (2) oral administration, and (3) topical application (e.g., eye drops, intranasal application, and intravaginal application).However, in a preferred embodiment, the composition is formulated for oral administration.

[0194] In some embodiments, the preferred route of administration of the compositions provided herein is oral. In such cases, the compositions for oral use have the manner, shape, or formulation described for the compositions of the invention in the fourth aspect of the invention.

[0195] In certain embodiments, the definitions and certain embodiments of the first, second, third, fourth and fifth aspects of the invention apply to the sixth aspect of the invention.

[0196] 5. Food and Nutritional Supplements In a seventh aspect, the present invention relates to a food supplement or dietary supplement comprising the composition of the fourth aspect of the invention or the composition obtainable by the method of the fifth aspect of the invention and a nutritionally acceptable excipient.

[0197] As used in the present invention, the term "food supplement" or "dietary supplement" refers to a concentrated source of nutrients or other substances obtained from an edible product, the purpose of which is to supplement the normal diet. Food supplements or dietary supplements according to the present invention include functional food compositions, i.e., foods, beverages, pet food or feed, or supplementary foods, beverages, pet food or feed, dietary supplements, flavors or flavorings, environmentally friendly or cosmetic formulations. In general, the term "food supplement" or "dietary supplement" may also mean: (i) A product intended to supplement the diet and having or containing one or more of the following nutritional ingredients: [A] vitamins, [B] minerals, [C] herbs or other botanical elements, [D] amino acids, or [E] nutritional substances for use by humans to supplement the diet by increasing total nutrient intake; or (F) a concentrate, metabolite, component, extract, or combination of any of the ingredients described in subparagraphs (A), (B), (C), (D), or (E); or (ii) Products that: (A) are intended to be ingested; (B) are not conventionally used as a food or as an independent product of a meal or the diet; and (C) are labeled as dietary supplements.

[0198] The "food supplement" or "nutraceutical" according to the present invention is usually administered orally and is provided together with the subject's diet. It can be in completely different forms, such as tablets, capsules, suspensions, dry powders, wet compositions, dry tube feeding, or wet tube feeding. It can be provided as a nutritional preparation, for example in the form of a medical food, for example in the form of tube feeding, or in the form of oral feeding as a complete meal, as a part of a meal, as a food additive, as a powder for a solution, for example a powder for a health drink, as a solution, as a prepared drink (such as juice, shake, yogurt drink, smoothie, or soy milk drink), as a bar, or dispersed in any type of food, such as baked goods, cereal bars, dairy bars, fast food, soup, breakfast cereal, muesli, candy, cookies, cake, etc.

[0199] As used herein, the term "nutritional acceptable excipient" or "nutritional acceptable supplement" refers to a supplement as defined above that is edible and is involved in the preparation of a solution to be administered orally. Typical nutritionally acceptable supplements, diluents, and excipients will be known to those skilled in the art. Non-limiting examples of such supplements are provided in U.S. Patent Nos. 6,258,846, 6,576,666, and 7,112,609.

[0200] The amount of the composition present in a human or animal dietary supplement composition, nutritional product, or food product (e.g., a functional food composition, i.e., a food, beverage, pet food or feed, or a supplement, beverage, pet food or feed), dietary supplement, flavor or flavoring agent, drug (pharmaceutical composition or formulation), veterinary composition, environmentally friendly, or cosmetic formulation will vary depending on the application. Typically, the amount of the composition present in a supplement or dietary supplement will be from about 0.001 percent to about 50 percent, e.g., from about 0.01 percent to about 10 percent, or from about 0.1 percent to 1 percent, by weight, of the dietary supplement composition, nutritional product, or food product, dietary supplement, flavor or flavoring agent, environmentally friendly formulation.

[0201] In certain embodiments, the definitions and certain embodiments of the first, second, third, fourth, fifth and sixth aspects of the invention apply to the seventh aspect of the invention.

[0202] 6. Use of the present invention as a medicine In an eighth aspect, the present invention relates to a composition or a kit of parts according to the fourth aspect of the invention, a composition obtainable by the method according to the fifth aspect of the invention, a pharmaceutical composition according to the sixth aspect of the invention, or a food supplement or dietary supplement according to the seventh aspect of the invention, for use in medicine.

[0203] In a ninth aspect, the present invention relates to a composition or a kit of parts according to the fourth aspect of the invention, a composition obtainable by the method according to the fifth aspect of the invention, a pharmaceutical composition according to the sixth aspect of the invention, or a food supplement or dietary supplement according to the seventh aspect of the invention, for use in the treatment of cardiovascular disease.

[0204] As used herein, the term "cardiovascular disease" or "CVD" refers to a disease that affects the heart and / or blood vessels. CVD includes coronary artery disease (CAD), such as angina pectoris and myocardial infarction. Other CVD includes atherosclerosis, stroke, heart failure, hypertensive heart disease, rheumatic heart disease, cardiomyopathy, arrhythmia, congenital heart disease, valvular heart disease, carditis, aortic aneurysm, peripheral artery disease, thromboembolic disease, and venous thrombosis. Peripheral vascular disease, cerebrovascular accident, or coronary artery disease is associated with atherosclerosis and generally involves the presence of atherosclerosis.

[0205] In certain embodiments, the cardiovascular disease of the ninth aspect of the invention is selected from the group consisting of atherosclerosis, cerebrovascular accidents, peripheral vascular disease, and coronary disease.

[0206] The term "atherosclerosis" is defined in the first, second or third aspect of the present invention.

[0207] As used herein, the term "peripheral vascular disease" or "PVD" refers to the blockage of a large artery that is not part of the coronary arteries, the aortic arch, or the cerebral vasculature. This causes acute or chronic ischemia (lack of blood supply). EVP may be causally related to atherosclerosis, an inflammatory process that leads to stenosis, embolism, or thrombus formation. In general, the term EVP is used to refer to atherosclerotic blockage in the lower extremities.

[0208] In a particular embodiment of the fourth aspect of the invention, the peripheral vascular disease is due to the presence of atherosclerosis.

[0209] As used herein, the terms "cerebrovascular accident", "brain attack", "stroke" or "cerebral infarction" refer to a situation in which blood flow to a part of the brain is stopped, causing cell death. An "ischemic cerebrovascular accident" or "ischemic stroke" is due to the absence of blood supply, which is different from a "hemorrhagic cerebrovascular accident" or "ischemic stroke" which is due to bleeding. As used herein, the terms "ischemic stroke" or "ischemic cardiovascular attack" refer to a situation in which cerebral structures lose their blood supply due to a sudden and immediate interruption of blood flow due to an occlusion of one of the arteries that supply the brain body. The occlusion may be due to fibrin or calcium accumulation, or red blood cell abnormalities, but generally is due to atherosclerosis or an embolus coming from another location, essentially the heart or another artery (e.g., the carotid artery or the bifurcation of the aortic arch) (cerebral embolism). As used herein, the term "hemorrhagic stroke" or "hemorrhagic cerebral attack" refers to a situation in which a cerebral blood vessel ruptures, causing areas of the brain that depend on this vessel, typically an artery, to lose their blood supply. The extravasation, along with other blood vessels, can put pressure on cerebral structures, thereby amplifying the affected area of ​​the brain.

[0210] In a particular embodiment of the ninth aspect of the invention, the cerebrovascular accident is ischemic, preferably due to the presence of atherosclerosis.

[0211] As used in the present invention, the term "coronary disease" or "coronary artery disease" refers to a situation in which there is an imbalance between coronary blood flow, i.e., coronary blood flow, and the oxygen requirement of the heart muscle, i.e., the myocardium. This imbalance causes ischemia, the effects of which are metabolic (elevated lactate, acidosis, decreased ATP, decreased phosphocreatine), mechanical (decreased cardiac contractility, decreased compliance of the ischemic area, etc.), and electrical (altered resting and action potentials, electrical instability, and secondary heart rhythm disorders). The main cause of coronary disease is narrowing of the coronary arteries that supply blood to the heart, due to atherosclerosis.

[0212] In a particular embodiment of the fourth aspect of the invention, the coronary disease is due to the presence of atherosclerosis.

[0213] Depending on the disorder and subject to be treated, and the route of administration, the compositions, components of the kit of parts, pharmaceutical compositions, food supplements, or dietary supplements of the present invention can be administered in various doses (i.e., administered to a patient in need thereof at a therapeutically effective dose). In this regard, as will be appreciated by those skilled in the art, the dose administered to a mammal, particularly a human, must be sufficient to provide a therapeutic effect in the mammal for a reasonable time frame in the context of the present invention. In certain embodiments, the therapeutic effect corresponds to the efficacy in treating a disease or condition described herein, as specified in the sixth aspect of the present invention. As will be appreciated by those skilled in the art, the choice of dose and precise composition, as well as the optimal dosing regimen, will also be influenced by the pharmacological properties of the formulation, the nature and severity of the condition to be treated, and the physical state of the receptor, as well as the age, condition, weight, sex, and response of the patient to be treated, and the stage / severity of the disease.

[0214] As used herein, the term "subject" refers to a mammal, preferably a human.

[0215] The subject to be treated is a mammal, preferably a human. The subject to be treated according to the present invention can be selected based on a number of criteria related to cardiovascular disease, such as serum oxLDL and / or LDL levels, or a corresponding oxLDL / LDL-cholesterol ratio.

[0216] In a particular embodiment, the subject presents with hypercholesterolemia, preferably moderate hypercholesterolemia, hi another particular embodiment, the subject presents with acute hypercholesterolemia.

[0217] Thus, in a preferred embodiment, the composition, kit of parts, pharmaceutical composition, or food or dietary supplement used in the eighth or ninth aspect of the invention is administered to a patient with hypercholesterolemia, preferably moderate hypercholesterolemia.

[0218] As used herein, the term "hypercholesterolemia" refers to a pathological condition characterized by high blood cholesterol levels. Considering that cholesterol is insoluble in water, it is transported by combining with proteins in plasma to form lipoproteins. Lipoproteins are classified according to their density into very low density lipoproteins (VLDL), intermediate density lipoproteins (IDL), low density lipoproteins (LDL), and high density lipoproteins (HDL). All lipoproteins transport cholesterol, but lipoproteins other than HDL, especially LDL-transport cholesterol (LDL-cholesterol), tend to deposit in arterial walls when their levels are high, which leads to the formation of atherosclerosis and coronary disease, stroke, and peripheral arterial disease. In general, cholesterol levels are considered high when the concentration of whole blood cholesterol is greater than 200 mg / dl and / or the concentration of LDL-cholesterol is greater than 100 mg / dl.

[0219] As used herein, the term "moderate hypercholesterolemia" refers to hypercholesterolemia characterized by a whole blood cholesterol concentration of more than 200 mg / dl and less than 250 mg / dl and an LDL-cholesterol concentration of more than 100 mg / dl and less than 175 mg / dl. As used herein, the term "acute hypercholesterolemia" refers to hypercholesterolemia characterized by a whole blood cholesterol concentration of more than 250 mg / dl and an LDL-cholesterol concentration of more than 175 mg / dl.

[0220] While the compositions, components of a kit of parts, pharmaceutical compositions, or supplements or dietary supplements of the invention can be administered by themselves, the present invention also contemplates that at least one of the components of the composition, kit of parts, pharmaceutical composition, or supplement or dietary supplement may be administered separately from the remainder of the components of the composition, kit of parts, pharmaceutical composition, or supplement or dietary supplement.

[0221] Thus, in certain embodiments of the eighth and ninth aspects of the invention, hydroxytyrosol and the polyphenol other than hydroxytyrosol are administered separately.

[0222] In another specific embodiment, hydroxytyrosol is administered separately from at least one polyphenol other than hydroxytyrosol or from an extract comprising at least one polyphenol other than hydroxytyrosol.In another specific embodiment, hydroxytyrosol is administered separately from at least one polyphenol other than hydroxytyrosol or from an extract comprising at least one polyphenol other than hydroxytyrosol and from the remainder of the components of the composition, kit of parts, pharmaceutical composition, or food or dietary supplement of the present invention.

[0223] In another embodiment, the plant extract containing hydroxytyrosol is administered separately from at least one polyphenol other than hydroxytyrosol or separately from the extract containing at least one polyphenol other than hydroxytyrosol. In another particular embodiment, the plant extract containing hydroxytyrosol is administered separately from at least one polyphenol other than hydroxytyrosol or separately from the extract containing at least one polyphenol other than hydroxytyrosol and from the remainder of the components of the composition, kit of parts, pharmaceutical composition, or food or dietary supplement of the invention.

[0224] In another particular embodiment, at least one polyphenol other than hydroxytyrosol is administered separately from hydroxytyrosol or from the extract containing hydroxytyrosol.In another particular embodiment, at least one polyphenol other than hydroxytyrosol is administered separately from hydroxytyrosol or from the extract containing hydroxytyrosol and from the remainder of the components of the composition, kit of parts, pharmaceutical composition, or food or dietary supplement of the invention.

[0225] In another particular embodiment, the plant extract comprising at least one polyphenol other than hydroxytyrosol is administered separately from hydroxytyrosol or from the extract comprising hydroxytyrosol.In another particular embodiment, the plant extract comprising at least one polyphenol other than hydroxytyrosol is administered separately from hydroxytyrosol or from the extract comprising hydroxytyrosol and from the remainder of the components of the composition, kit of parts, pharmaceutical composition, or food or dietary supplement of the invention.

[0226] In certain embodiments, the different parts of the kit used in the eighth and ninth aspects of the invention can be administered separately or can be combined together before administration. In preferred embodiments of the eighth and ninth aspects of the invention, the components of the kit of parts are combined together before administration.

[0227] In an embodiment of the eighth and ninth aspects of the invention, administering the composition, component of the kit of parts, pharmaceutical product, or food or dietary supplement comprises administering the composition, component of the kit of parts, pharmaceutical product, or food or dietary supplement of the fourth aspect of the invention in multiple doses for at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 1 week, at least 1.5 weeks, at least 2 weeks, at least 2.5 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, at least 6 weeks, at least 7 weeks, at least 8 weeks, at least 9 weeks, at least 10 weeks, at least 11 weeks, at least 12 weeks, preferably at least 1 week, more preferably at least 2 weeks, even more preferably at least 4 weeks, or at least 8 weeks.

[0228] In another embodiment, administration of the composition, component of a kit of parts, pharmaceutical product, or food or dietary supplement comprises administering a daily dose of the composition, component of a kit of parts, pharmaceutical product, or food or dietary supplement of the invention 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 15, 20, preferably 2, times.

[0229] As those skilled in the art will understand, the initial setting of suitable therapeutic dose of the composition of the present invention can be identified in vitro and in vivo animal model systems, as well as in human clinical trials. Those skilled in the art will know how to utilize animal experiments and human experience to identify doses that can be safely administered without causing toxicity or other side effects. For acute treatments where it is desired to substantially restore serum oxLDL levels and serum oxLDL / LDL ratios, therapeutic doses are preferably close to the maximum tolerated dose. For chronic preventative use, lower doses may be desirable due to concerns about long-term effects.

[0230] In another embodiment of the eighth and ninth aspects of the invention, the administration of the composition, the component of the kit of parts, the pharmaceutical composition, or the food or dietary supplement provides hydroxytyrosol at 1 mg / day, 2 mg / day, 3 mg / day, 4 mg / day, 5 mg / day, 6 mg / day, 7 mg / day, 7.5 mg / day, 8 mg / day, 8.5 mg / day, 9 mg / day, 9.5 mg / day, 10 mg / day, 10.5 mg / day, 11 mg / day, 11.5 mg / day, 12 mg / day, 13 mg / day, 14 mg / day, 15 mg / day, 17 mg / day, 20 mg / day, 22 mg / day, 25 mg / day, 30 mg / day, 35 mg / day, 40 mg / day, 45 mg / day, 50 mg / day, 60 mg / day, 70 mg / day, 80 mg / day, 90 mg / day, 100 mg / day, 120 mg / day, 120 mg / day, 120 mg / day, 120 mg / day, 120 mg / day, mg / day, preferably 7.5 mg / day.

[0231] In another particular embodiment of the eighth and ninth aspects of the invention, the administration of the composition, the component of the kit of parts, the pharmaceutical composition, or the food or dietary supplement comprises administering a polyphenol other than hydroxytyrosol at 20 mg / day, 30 mg / day, 40 mg / day, 50 mg / day, 60 mg / day, 70 mg / day, 80 mg / day, 90 mg / day, 100 mg / day, 125 mg / day, 150 mg / day, 175 mg / day, 190 mg / day, 200 mg / day, 205 mg / day, 210 mg / day, 215 mg / day, 220 mg / day, 230 mg / day, 240 mg / day, 250 mg / day, 275 mg / day, 300 mg / day, 325 mg / day, 350 mg / day, 375 mg / day, 400 mg / day, preferably 210 mg / day.

[0232] In another particular embodiment of the eighth and ninth aspects of the invention, the administration of the composition, the component of the kit of parts, the pharmaceutical composition, or the food or dietary supplement provides hydroxytyrosol at 1 mg / day, 2 mg / day, 3 mg / day, 4 mg / day, 5 mg / day, 6 mg / day, 7 mg / day, 7.5 mg / day, 8 mg / day, 8.5 mg / day, 9 mg / day, 9.5 mg / day, 10 mg / day, 10.5 mg / day, 11 mg / day, 11.5 mg / day, 12 mg / day, 13 mg / day, 14 mg / day, 15 mg / day, 17 mg / day, 20 mg / day, 22 mg / day, 25 mg / day, 30 mg / day, 35 mg / day, 40 mg / day, 45 mg / day, 50 mg / day, 60 mg / day, 70 mg / day, 80 mg / day, 90 mg / day, 100 mg / day, 120 mg / day, 120 mg / day, 120 mg / day, 120 mg / day, 120 mg / day, 120 mg / day, 120 mg / day, 120 mg / day, mg / day, preferably 7.5 mg / day, and polyphenols other than hydroxytyrosol at any of the mg / day doses indicated above, preferably 210 mg / day of polyphenols other than hydroxytyrosol.

[0233] Thus, in certain embodiments of the eighth and ninth aspects of the invention, administration of the composition, component of the kit of parts, pharmaceutical product, or food or dietary supplement of the invention comprises administration of 7.5 mg / day of hydroxytyrosol and 210 mg / day of a polyphenol other than hydroxytyrosol.

[0234] In another particular embodiment of the eighth and ninth aspects of the invention, the administration of the composition, the component of the kit of parts, the pharmaceutical composition, or the food or dietary supplement comprises administering at least one of the following: 10 mg / day, 15 mg / day, 20 mg / day, 25 mg / day, 30 mg / day, 35 mg / day, 40 mg / day, 45 mg / day, 50 mg / day, 55 mg / day, 57 mg / day, 60 mg / day, 61 mg / day, 62 mg / day, 63 mg / day, 64 mg / day, 65 mg / day, 66 mg / day, 67 mg / day, 68 mg / day, 69 mg / day, 70 mg / day, 72 mg / day, 75 mg / day, 77 mg / day, 80 mg / day, 82 mg / day, 85 mg / day, 87 mg / day, 90 mg / day, 95 mg / day, ... mg / day, 100 mg / day, 110 mg / day, 120 mg / day, 150 mg / day, 170 mg / day, 200 mg / day, 225 mg / day, 250 mg / day, 275 mg / day, 300 mg / day, preferably 67 mg / day.

[0235] In another particular embodiment of the eighth and ninth aspects of the invention, the administration of the composition, the component of the kit of parts, the pharmaceutical composition, or the food or dietary supplement comprises administering at least 50 mg / day, 70 mg / day, 100 mg / day, 125 mg / day, 150 mg / day, 175 mg / day, 200 mg / day, 225 mg / day, 250 mg / day, 275 mg / day, 300 mg / day, 325 mg / day, 350 mg / day, 375 mg / day, 400 mg / day, 425 mg / day, 450 mg / day, 475 mg / day, 500 mg / day, 525 mg / day, 550 mg / day, 575 mg / day, 600 mg / day, 625 mg / day, 650 mg / day, 675 mg / day, 680 mg / day, 690 mg / day, 70 mg / day, 70 mg / day, 80 mg / day, 85 mg / day, 90 mg / day, 95 mg / day, 100 mg / day, 125 mg / day, 150 mg / day, 175 mg / day, 200 mg / day, 225 mg / day, 250 mg / day, 275 mg / day, 300 mg / day, 325 mg / day, 350 mg / day, 375 mg / day, 400 mg / day, 425 mg / day, 450 mg / day, 475 mg / day, 500 mg / day, 525 mg / day, 550 mg / day, 575 mg / day, 600 mg / day, 625 mg / day, 650 mg / day, 675 mg / day, 680 mg / day, mg / day, 700 mg / day, 710 mg / day, 720 mg / day, 730 mg / day, 740 mg / day, 750 mg / day, 775 mg / day, 800 mg / day, 825 mg / day, 850 mg / day, 875 mg / day, 900 mg / day, 950 mg / day, 1 g / day, 2.5 g / day, 3 g / day, 3.5 g / day, 4 g / day, 5 g / day, 6 g / day, 7 g / day, 8 g / day, 9 g / day, 10 g / day, preferably 700 mg / day.

[0236] In another particular embodiment of the eighth and ninth aspects of the invention, the administration of the composition, the component of the kit of parts, the pharmaceutical composition, or the food or dietary supplement comprises administering at least one of the following: 10 mg / day, 15 mg / day, 20 mg / day, 25 mg / day, 30 mg / day, 35 mg / day, 40 mg / day, 45 mg / day, 50 mg / day, 55 mg / day, 57 mg / day, 60 mg / day, 61 mg / day, 62 mg / day, 63 mg / day, 64 mg / day, 65 mg / day, 66 mg / day, 67 mg / day, 68 mg / day, 69 mg / day, 70 mg / day, 72 mg / day, 75 mg / day, 77 mg / day, 80 mg / day, 82 mg / day, 85 mg / day, 87 mg / day, 90 mg / day, 95 mg / day, ... mg / day, 100 mg / day, 110 mg / day, 120 mg / day, 150 mg / day, 170 mg / day, 200 mg / day, 225 mg / day, 250 mg / day, 275 mg / day, 300 mg / day, preferably 67 mg / day, and administering a plant extract providing polyphenols as defined in the first to fourth aspects of the invention at any of the mg / day doses indicated above, preferably 700 mg / day.

[0237] In a preferred embodiment of the eighth and ninth aspects of the invention, administration of the composition, the component of the kit of parts, the pharmaceutical composition or the food or dietary supplement comprises administration of 67 mg / day of a plant extract providing hydroxytyrosol as defined in the first to fourth aspects of the invention and 700 mg / day of a plant extract providing polyphenols according to the first to fourth aspects of the invention.

[0238] In certain embodiments, the definitions and certain embodiments of the first, second, third, fourth, fifth, sixth and seventh aspects of the invention apply to the eighth and ninth aspects of the invention.

[0239] In certain embodiments, the terms "consists of," "essentially consists of," and "comprises" are synonymous in any embodiment of any aspect of the present invention.

[0240] The invention will now be described by way of examples which are merely illustrative and not limiting of the scope of the invention. EXAMPLES

[0241] Materials and Methods A. Materials and Methods Corresponding to Examples 1 and 2 Test subjects Volunteers were recruited at the Endocrinology Department of San Cecilio Hospital, Granada (Spain) and from the database of the Clinical Trials Department of Biosearch, SA. Participants in the study fulfilled the following inclusion criteria: male or female, aged 18-65 years, with moderate hypercholesterolemia (LDL-col > 100 mg / dL and / or total cholesterol > 200 mg / dL) and no pharmacological treatment for this condition. Exclusion criteria included pregnancy, significant medical problems, diabetes, and / or neurovascular disease, and allergies to antibiotics or any of the study components. Subjects were also excluded if they were following a dietary regimen or taking supplements during the study. The study was performed in accordance with the Declaration of Helsinki and the protocol was approved by the local Ethics Committee (Granada, Spain). Informed consent was obtained from all subjects. The clinical trial was registered in the US Library of Medicine (http: / / www.clinicaltrial.gov) under NCT04029727.

[0242] Approximately 300 subjects were contacted and invited to participate in the study. The suitability of 63 candidates was assessed. Of these candidates, 31 were excluded (11 did not meet the inclusion criteria and the rest refused to participate). Finally, a total of 32 subjects agreed to participate in the treatment intervention. These 32 subjects were randomly assigned to the study group (Figure 1).

[0243] Test Plan The study was a randomized, parallel-group, double-blind, placebo-controlled pilot study of 8 weeks duration. The study took place from May to July 2019. Thirty-two subjects were recruited and randomly assigned to two different groups according to a randomization scheme generated by the SIGESMU™ program. One group (extract group) received an oral supplement with a combination of olive fruit extract and almond skin extract (HT 7.5 mg + ASP 210 mg + maltodextrin 33 mg per day), while the other group (control group) received a placebo (maltodextrin 800 mg per day). The treatment lasted for 8 weeks, during which the subjects took two capsules per day with lunch (supplement containing the extract or placebo). Participants were instructed not to deviate from their regular habits and to maintain their usual diet and exercise regime for the 8 weeks. Neither the researchers nor the subjects knew the treatment sequence to which the subjects were assigned. The researchers were blinded to this information until the end of the study.

[0244] Test item Each capsule containing the extract contained 3.75 mg of HT in 33.5 mg of olive tree (Olea europaea L.) extract, 105 mg of ASP in 350 mg of almond skin (Prunus dulcis (Mill.)) extract, and 16.5 mg of maltodextrin. Each capsule containing the placebo contained 400 mg of maltodextrin. Both extracts were obtained from Biosearch, SA production plant in Talayuela, Caceres (Spain) and the blending of the extracts was carried out at Biosearch, SA R&D facility in Granada (Spain). The capsules were prepared at the Department of Pharmaceutical Technology, School of Pharmacy, University of Granada (Spain). The extract and the placebo were provided in identical gelatin capsules and packed in identical plastic containers. The containers were coded with a code number indicating the volunteer number according to the randomization.

[0245] Preparation of plant extracts The olive tree extract can be obtained by a method based on extraction of the leaves of O. europaea L. with an aqueous solvent, such as water. This method comprises: (i) extracting from the leaves of O. europaea using an aqueous solvent at a temperature of about 40° C. to 100° C. for 1 hour to 10 hours; (ii) treating the extract obtained in step (i) with activated carbon and then removing the activated carbon; Includes.

[0246] An extract of Prunus dulcis Mill. can be obtained following the same method used to obtain an extract of O. europaea L., except that the starting material is the pericarp instead of the leaves of O. europaea.

[0247] Optionally, step (i) may include multiple extraction steps using the initial solvent or different solvents to extract from the plant product, and the resulting extracts are combined and continued in the process. Alternatively, the extract may be concentrated by chromatography using a polymeric adsorbent resin. The method may also include a final drying step to dry the extract obtained in the above steps of the method.

[0248] Test parameters and data acquisition The primary outcome of the study was the determination of oxLDL levels.Secondary parameters included the oxLDL / LDL-c ratio, total cholesterol (TC), LDL cholesterol (LDL-c), HDL cholesterol (HDL-c) levels, and serum triglyceride (TG) levels.

[0249] Subjects were present at the Biosearch, SA facility in Granada at the start of the study and at weeks 4 and 8. Blood samples were taken before the start of the intervention and at each follow-up visit, after an overnight fast of at least 10 hours, using the Vacutainer™ SST™ II Advance Tubes (BD, NJ, USA) system containing thixotropic gel. Blood samples were centrifuged at 1000 g for 15 minutes to obtain serum, which was stored at -80°C.

[0250] Anthropometric measurements were performed at each visit using standard methods. Weight was measured using a Tanita BC-418 body composition analyzer (Tanita Corporation, Tokyo, Japan). Height was measured using a height measuring device with 1 mm accuracy (range, 80 cm–200 cm). Body mass index (BMI) was calculated as weight / height squared (kg / m 2 ) All measurements were performed by qualified personnel.

[0251] To control for possible changes in subjects' habits, each subject's food intake was assessed at the beginning and end of the study. Subjects recorded a full 72-hour breakdown of their dietary intake, including the type of food consumed and the weight of each meal. Daily diet, energy intake, nutrient intake, and energy provided by macronutrients were calculated using the Nutriber computer application (Funiber, Barcelona, ​​Spain). Subjects were also asked about their physical activity habits (type of activity and the amount of time the subject spent per week) at the beginning and end of the study.

[0252] Compliance with the protocol regarding consumption of the test product was verified at the end of the intervention by comparing the number of capsules supplied with the number of capsules returned. Adverse events were defined as any unfavourable or undesirable effect and were recorded at follow-up visits (4 and 8 weeks).

[0253] Analysis of biochemical parameters Levels of fluid parameters (TC, LDL-c, HDL-c, and TG) were analyzed by standard methods by an external laboratory (Reference Laboratory SA, Barcelona, ​​Spain). Serum oxLDL concentrations were determined by a specific ELISA for human oxLDL (Elabscience Biotechnology, USA). The oxLDL / LDL-c ratio was calculated.

[0254] statistical analysis Normal distribution of all variables was verified by normal probability graphs and the Shapiro-Wilk test. Data are expressed as mean (and SD) for continuous variables and n (%) for categorical variables.

[0255] For comparisons between groups (sampled vs. control) at the start of the study, continuous variables were analyzed with Student's t-test or Kruskal-Wallis nonparametric methods as appropriate, and categorical variables were analyzed with chi-squared tests.

[0256] All parameters were compared between groups and the variability with respect to the initial within-group values ​​was also analyzed. For that purpose, fitting tests based on bivariate analysis and mixed regression models will be performed. Comparisons between the experimental and control group means were performed by T-tests when it was possible to assume normality. When it was not possible to assume normality, the nonparametric Mann-Whitney U-test was performed. Furthermore, mixed linear regression models representing repeated measurements throughout the study (within-subject random effects) were fitted to the responses in order to examine the changes over time and between groups, as well as to find significant factors associated with the response.

[0257] A common alpha level of 0.05 was used as the cutoff point for statistical significance. Statistical analyses were performed using SPSS version 26.0 software for Windows (SPSS, Chicago, IL, USA).

[0258] B. Materials and Methods Corresponding to Examples 3 and 4 reagent α-Amylase, from porcine pancreas (Sigma-Aldrich A3176 or equivalent) Pepsin, from porcine gastric mucosa (Sigma-Aldrich P7000 or equivalent) Trypsin, from bovine pancreas (Sigma-Aldrich T8003 or equivalent) Pancreatin, from porcine pancreas (Sigma-Aldrich P1750 or equivalent) Bile salts (Sigma-Aldrich B8631 or equivalent) Basal medium (''Medium used to feed the reactor SHIME'' in Molly K. et al. Microbial Ecology in Health and Disease 7:191-200)

[0259] Table 1. Plant extracts used in this study plant extract Extracts Characterized active ingredients Polyphenol types Olive Tree Extract Hydroxytyrosol Flavonoids Almond Extract Total Polyphenols Flavonoids Grapefruit extract Flavonoids Flavonoids Flax extract Lignan Lignan Polygonum extract Resveratrol Stilbene [Table 1]

[0260] The olive wood and almond extracts correspond to those used in Materials and Methods, Section A. They can therefore also be obtained using the methods set out in that section.

[0261] The grapefruit extract, the flax extract and the knotweed extract can be obtained by applying the same method as that for obtaining the olive tree extract, indicated in section A of the Materials and Methods, in which: The starting material for grapefruit extract is the seed of the fruit of the Citrus Paradisi MacFad tree. The starting material for the flax extract is the seed of the fruit of Linum usitatissimum L. The starting material for the Polygonum cuspidatum extract is the root of Polygonum cuspidatum Sieb. et Zucc.

[0262] Furthermore, the solvent used in the processes for obtaining these extracts is a hydroalcoholic alcohol, which can be, for example, ethanol or methanol.

[0263] Chromatographic analysis The solutions of each extract were separated by chromatography on an Agilent 1200s instrument equipped with a binary pump and a DAD SL detector, using Phenomenex Luna C18 5 μm 250×4.6 mm as stationary phase at 30° C. and A: 1% formic acid (v / v); B: acetonitrile:methanol 80:20 (flow rate: 0.8 mL / min to 1 mL / min) as mobile phase. 10 μL of sample was injected and absorbance at 330 nm and 280 nm was measured during the run (65 min). The chromatographic conditions in terms of mobile phase composition and flow rate are summarized in the table below.

[0264] Table 2. Chromatographic conditions [Table 2] Time (min) Mobile phase flow rate (mL / min)

[0265] Small scale in vitro digestion A small-scale simulation of human (adult) digestion was performed at room temperature in four semi-series connected compartments, each with the characteristics shown in Table 3.

[0266] Table 3. Small-scale compartments simulating digestion processes [Table 3] Section Time (min) Volume Composition initial maximum Oral: Extract of saline solution α-amylase 50 U / mL Stomach: Medium B Pepsin 50 μg / mL Small intestine medium B Trypsin 394 μg / mL Pancreatin 250 μg / mL Bile salts 590 μg / mL Colon Medium B Human colonic microbiota

[0267] The steps of the digestion process and the transport of samples between compartments for treatment or to simulate a dynamic process are summarized in Table 4.

[0268] Table 4. Small scale digestion process. [Table 4] Time 1 stomach 1 small intestine 1 colon 1 1 The volume (mL) contained in each compartment at any given time (min), and 2 The volumes transported between compartments are shown: MS, mouth to stomach; S-SI, stomach to small intestine; SI-C, small intestine to colon. 3 Volumes of acid and base added to the stomach and small intestine, respectively, to change the pH from 4 (early stomach) to 2 (final stomach) and to maintain the pH value near 6.5 (small intestine). The time intervals during which digestion is occurring in each compartment are highlighted in bold and underlined.

[0269] Example 1. Effect of a combination of standardized olive tree (Olea europaea L.) extract and standardized almond (Prunus dulcis Mill.) skin extract on serum oxLDL levels Two subjects in the extract group refused to participate before attending the initial visit, leaving a total of 30 volunteers who completed the study (Figure 1). The compliance rate was found to be extremely high (approximately 100%). No adverse events due to the intake of the treatment capsules were reported for either of the two capsules.

[0270] Of the 30 volunteers who completed the study, 13 (43.43%) were men and 17 (56.7%) were women. The mean age of the subjects was 48.13 ± 12.9 years, and the mean BMI of the subjects was 25.38 ± 3.33 kg / m 2 No significant differences were detected in the baseline characteristics of the volunteers (Table 5).

[0271] Table 5. Initial characteristics of subjects enrolled in the study [Table 5] Control group Extract group Intragroup P Age (years) sex male woman Obesity / Overweight Smokers Participate in regular physical activity Total cholesterol LDL-cholesterol HDL-cholesterol Triglycerides Oxidized LDL Oxidized LDL / LDL-c ratio Continuous variables are mean ± standard deviation (SD), categorical variables are n (%). P indicates difference between groups.

[0272] Table 6 shows the energy and nutrient intake at baseline and at week 8. There was no significant difference between the intervention groups at baseline, and no significant difference between baseline and the end of the intervention in each group (extract or control). There was no change in the level of physical activity during the study (data not shown), and there was no significant change in BMI between the start and week 8 in both groups (Table 6).

[0273] Table 6. Dietary and anthropometric parameters of subjects receiving placebo or extract for 8 weeks. [Table 6] Control group Extract group Initial period 8th week P section Initial period 8th week P section Energy full fat saturated type Monosaturated polysaturated type Dietary Cholesterol protein carbohydrates Dietary fiber (g / day) Mean ± SD. P indicates the difference between the initial value and the value after 8 weeks of treatment in each group. %En: Values ​​expressed as % of total energy.

[0274] Table 7 shows the values ​​of lipid parameters and oxidized lipid variables evaluated in the study. No significant changes were observed between baseline and the end of the intervention in serum TC, LDL-c, HDL-c, and TG in any group. Regarding lipid oxidation status, oxLDL levels were significantly increased from baseline in the control group at 4 weeks (p=0.01) and 8 weeks (p=0.03), whereas in the extract group, oxLDL levels tended to decrease and were significantly lower in the extract group than in the control group after 8 weeks of treatment (p<0.001). When analyzing the oxLDL / LDL-c ratio, an accurate estimate of the oxidation of LDL (19) in vivo, a significant increase from baseline in the oxLDL / LDL-c ratio was observed in the control group at 4 weeks (p=0.001) and 8 weeks (p=0.002), whereas in the extract group, a significant decrease in this ratio was observed at 8 weeks (p=0.047). The levels of oxLDL / LDL-c ratio in the extract group were also significantly lower than those in the control group after 8 weeks of treatment (p<0.001) (Table 7 and Figure 2).

[0275] Table 7. Lipid and oxidized LDL parameters at the start of treatment, 4 weeks of treatment, and 8 weeks of treatment in subjects receiving placebo or the extract. [Table 7] Control group Extract group Intragroup P Total cholesterol initial stage Week 4 Week 8 LDL-cholesterol HDL-cholesterol Triglycerides OxLDL / LDL-c ratio Means ± SD adjusted for sex, age, obesity / overweight, smoking, and physical activity. P indicates difference between groups at each time point. * P<0.05 at baseline, 4 weeks, and 8 weeks

[0276] Example 2. Effect of a mixture of olive wood, almond, grapefruit, flax, and knotweed extracts on the bioavailability of hydroxytyrosol from olive wood extract To verify whether the presence of other polyphenols favors the bioavailability of hydroxytyrosol present in olive wood extract, small-scale digestion assays were carried out using the extracts separately and in various combinations. As a reference, 50 mg / mL olive wood extract was used, as well as its mixtures with almond skin extract at 1:1, 1:4, and 1:9. Based on the ratio of polyphenols of both extracts in each mixture, the mixtures with the other three extracts to be assayed were determined (Table 9).

[0277] Table 9. Mixtures of extracts, their initial concentration in the mouth (mg / mL) and polyphenol concentration (mg / mL). [Table 8] Almond extract polyphenols Mixture Olive Wood Almond Ratio HT Flavonoid Ratio Grapefruit extract polyphenols Mixture Olive Tree Grapefruit Ratio HT Flavonoid Ratio Flax Extract Polyphenols Mixture Olive Wood Flax Ratio HT Lignan Ratio Polygonum extract polyphenol Mixture Olive Wood Polygonum Ratio HT Stilbene Ratio

[0278] First, a mixture of olive wood and almond extracts was assayed and the hydroxytyrosol content of the final small intestine sample (digestion 370 min) derived from this mixture was analyzed by HPLC-DAD, followed by interpolation of the data obtained from the hydroxytyrosol calibration line. Table 10 shows the results of this analysis (ppm).

[0279] Table 10. HT concentrations (ppm) in small intestine and colon samples in vitro (small scale) and % of theoretical maximum derived from extract dose and dilution ratio. [Table 9] mixture

[0280] The results of this assay seem to show, first of all, that under the conditions of this assay, after the digestive process in a compartment simulating the small intestine, the HT present in the olive wood extract is degraded by about 50%. Furthermore, the presence of other polyphenols allows the concentration of HT to increase, which does so exponentially. Under these experimental conditions, the maximum concentration of almonds may allow close to 90% of the HT to be consumed and available for absorption in the intestinal lumen.

[0281] Considering that in vivo absorption of HT probably occurs in the small intestine, the presence of this phenolic acid was analyzed only in intestinal samples (370 min digestion) obtained in small-scale digestion assays performed with mixtures of olive wood extracts and other extracts (Table 11).

[0282] Table 11. HT concentrations (ppm) in in vitro small intestine samples (small scale) and % of theoretical maximum (340.2 ppm) derived from extract dose and dilution rate. [Table 10] Grapefruit Flax Polygonum mixture

[0283] The results obtained show that the presence of other polyphenols, regardless of their structure or origin, provides a degree of protection against the hydroxytyrosol of the olive wood extract, equivalent to that detected in the case of the polyphenols of the almond extract.

[0284] Example 4. Analysis of polyphenols present in each plant extract Regarding the characterization of plant extracts, polyphenol analysis methods by high performance liquid chromatography (HPLC) have been developed in-house by Biosearch, SA for the quantitative analysis of polyphenols in plant extracts and complex mixtures, allowing classification of polyphenols based on the retention times obtained, and in some cases by polyphenol family, or combined with previous information using the response of corresponding chromatographic standards for the quantitative analysis of specific polyphenols.

[0285] Although there are other methods for polyphenol identification by HPLC, the advantage of the method used is that it allows the separation of different families and types of polyphenols in the same chromatogram, thus reducing to the greatest extent possible the overlap between the signals, which may consequently interfere with the identification of the polyphenols. This efficiency in terms of signal separation is achieved as a result of the chromatographic conditions of the method (previously shown). Under these conditions, a chromatogram of 65 minutes is obtained, which, as previously shown, allows the different signals corresponding to the polyphenols of the sample to be separated without overlap between the different families of polyphenols. A DAD detector is used, set to detect signals with the usual wavelengths for the detection of polyphenols: 280 nm and 330 nm.

[0286] The chromatographic assay carried out allows the demonstration that hydroxytyrosol present in the olive tree fruit extract is not present in the other evaluated samples containing other types of polyphenols (peak with a retention time (RT) of about 10.5 min detected at 280 nm; Figures 3 to 6). Similarly, the main group of compounds with a RT greater than 35 min, detected both at 280 nm and at 330 nm in the almond extract, is not present in the olive tree extract and their presence in the components of this extract can be considered as residual (Figure 3). When the olive tree and grapefruit extracts are analyzed jointly, the results are similar: the main components of each extract (hydroxytyrosol with a RT of 10.5 min in olive tree and flavonoids with a RT between 35 and 45 min in grapefruit) are not detected in the other extract (Figure 4). Comparing the polyphenol composition of the olive tree and flax extracts again confirms that compounds with a RT greater than 40 min, such as lignans, are not detected in the majority of the olive tree extract (Figure 5). Finally, the Polygonum extract is characterized by the presence of a high concentration of the stilbene resveratrol, which has a RT of about 43 min, and the absence of compounds with a RT of 10-11 min (characteristic of hydroxytyrosol), a type of compound that is prominently detected in the olive wood extract in this chromatogram (Figure 6).

[0287] Based on these results, it can be affirmed that hydroxytyrosol present in the olive tree fruit extract is not present in the other evaluated samples containing other types of polyphenols, and similarly, the flavonoids, lignans and stilbenes that characterize almond extract, grapefruit extract, flax extract and polygonum extract are not found prominently in the olive tree extract. [Explanation of symbols]

[0288] Figure 1 Volunteers contacted (~~300) Selected Excluded Did not meet inclusion criteria Refused to participate Randomized Extract Group Dropouts Completed study Control Group Figure 2 oxLDL / LDL-cholesterol ratio (ng / mg) Baseline 4-Week 4th week 8-Week 8th week Control Extract Data collection points during intervention Figure 3 Olive tree Almond Figure 4 Olive tree Grapefruit Figure 5 Olive tree Flax Figure 6 Olive tree Polygonum knotweed< / n> < / n> < / n>

Claims

1. A pharmaceutical product or a food supplement or nutraceutical comprising almond skin polyphenol (ASP) for increasing the bioavailability of hydroxytyrosol after oral administration of hydroxytyrosol or for increasing the anti-atherosclerotic effect of hydroxytyrosol after oral administration of hydroxytyrosol, wherein hydroxytyrosol is part of an extract of the fruit of Olea europaea, and in an oral administration embodiment the w / w ratio of hydroxytyrosol to almond skin polyphenol (ASP) is 1:

28.

2. 2. A pharmaceutical product or a food or nutritional supplement according to claim 1 for use in the prevention and / or treatment of cardiovascular diseases.

3. 3. The pharmaceutical product or food or dietary supplement of claim 2, wherein the cardiovascular disease is selected from the group consisting of atherosclerosis, cerebrovascular accident, peripheral vascular disease, and coronary disease.

4. The pharmaceutical product or food or dietary supplement according to any one of claims 1 to 3, wherein the almond skin polyphenols (ASP) are administered orally.

5. 2. The pharmaceutical product or food or dietary supplement of claim 1, wherein the hydroxytyrosol-containing extract of the fruit of Olea europaea contains 1% to 90% w / w hydroxytyrosol.

6. 2. A pharmaceutical product or a food or dietary supplement as claimed in claim 1, wherein the hydroxytyrosol-containing extract of the fruit of Olea europaea contains 10% (w / w) hydroxytyrosol.

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