AU2021228197A1

AU2021228197A1 - Uses and compositions based on polyphenols for improving the oral bioavailability of hydroxytyrosol

Info

Publication number: AU2021228197A1
Application number: AU2021228197A
Authority: AU
Inventors: Óscar Bañuelos Hortigüela; Ruth Blanco Rojo; José Luis LÓPEZ LARRAMENDI; José Antonio MALDONADO LOBÓN; Mónica María OLIVARES MARTÍN; Luis PÉREZ MARTÍNEZ
Original assignee: Biosearch SA
Current assignee: Biosearch SA
Priority date: 2020-02-28
Filing date: 2021-02-26
Publication date: 2022-07-07
Also published as: TWI795736B; JP2023514680A; EP4110309A1; CO2022011987A2; BR112022015705A2; WO2021170806A1; MX2022010216A; CN115003291A; TW202131912A; KR20220123074A

Abstract

The invention relates to compositions of a natural origin, comprising hydroxytyrosol and polyphenols which are not hydroxytyrosol and do not occur naturally in the olive tree (O. europaea), characterized by allowing particularly high bioavailability of hydroxytyrosol and a particularly high antiatherosclerotic activity of hydroxytyrosol, once orally administered. Said compositions can consist of mixtures of plant extracts. The invention also relates to the use of said compositions for the treatment of cardiovascular diseases, particularly atherosclerosis.

Description

USES AND COMPOSITIONS BASED ON POLYPHENOLS FOR IMPROVING THE ORAL BIOAVAILABILITY OF HYDROXYTYROSOL

FIELD OF THE INVENTION

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

BACKGROUND OF THE INVENTION

Cardiovascular diseases (CVDs) are still the main cause of death worldwide and are responsible for about 17.8 million deaths in 2017, which represents a 12.5% increase in mortality over the last decade.

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 with endothelial dysfunction, in which it has been suggested that oxidized low-density lipoproteins (oxLDL) play an important role. oxLDLs bind with higher affinity than native LDLs do to several receptors, so their absorption by macrophages is more rapid, which leads to the buildup of cholesterol and to the formation of atherosclerotic plaque. Furthermore, oxLDLs promote endothelial cell activation and dysfunction, vascular smooth muscle cell migration and proliferation, and platelet activation, causing an inflammatory response, endothelial dysfunction; aggravating the atherogenic process. As a result, the concentration of circulating oxLDL is considered one of the most important biomarkers of the progression of atherosclerosis.

Generally, the treatment of atherosclerosis consists of changing the lifestyle to a healthier one. However, if this is not enough, it is usually combined with the administration of drugs which in most cases are targeted at reducing blood pressure and/or cholesterol. In advanced cases, surgical intervention may be necessary to dilate the narrowed artery/arteries. Therefore, there is a need in the state of the art for treatments that allow increasing the benefits of a healthy lifestyle over the development of atherosclerosis. SUMMARY OF THE INVENTION

The present invention is based in the finding that combinations of hydroxytyrosol (HT) present in olive oil with polyphenols not present naturally in the olive tree ( Olea europaea ) or in olive oil, such as those present in an almond skin extract, in a grapefruit, flax, or polygonum extract, allow increasing the bioavailability of HT after their oral administration in comparison with the administration of HT in the absence of polyphenols. Furthermore, the inventors have observed how the administration of a composition comprising HT and almond skin polyphenols (ASP) to subjects with moderate hypercholesterolemia (treated group) reduces serum levels of oxidized low-density lipoproteins (oxLDL), as well as the oxLDL/LDL ratio. However, in subjects with moderate hypercholesterolemia treated with placebo (control group), serum levels of oxLDL increase after the treatment, as does the oxLDL/LDL ratio.

Therefore, a first aspect of the invention relates to the use of at least one polyphenol that is not hydroxytyrosol and/or that does not occur naturally in the tree Olea europaea ((). europaea ) for increasing the bioavailability of hydroxytyrosol after the oral administration of hydroxytyrosol or for increasing the antiatherosclerotic effect of hydroxytyrosol after its oral administration.

In a second aspect, the invention relates to a polyphenol that is not hydroxytyrosol and/or that does not occur naturally in the tree O. europaea for use in increasing the bioavailability of hydroxytyrosol after the oral administration of hydroxytyrosol or in increasing the antiatherosclerotic activity of hydroxytyrosol after its oral administration.

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

In a fourth aspect, the invention relates to a composition or kit of parts comprising hydroxytyrosol and at least one polyphenol, wherein the polyphenol that is not hydroxytyrosol and/or wherein the at least one polyphenol does not occur naturally in the tree O. europaea. In a fifth aspect, the invention relates to a method for obtaining the composition of the fourth aspect of the invention, which method comprises contacting a composition comprising hydroxytyrosol with a composition comprising at least one polyphenol that is not hydroxytyrosol and/or that does not occur naturally in the tree (). europaea.

In a sixth aspect, the invention relates to a pharmaceutical composition comprising the composition of the fourth aspect of the invention, or a composition obtained by the method of the fifth aspect of the invention, and a pharmaceutically acceptable excipient.

In a seventh aspect, the invention relates to a food or nutritional supplement comprising the composition of the fourth aspect of the invention, or a composition obtained by the method of the fifth aspect of the invention, and a nutritionally acceptable excipient.

In an eighth aspect, the invention relates to the composition or kit of parts of the fourth aspect of the invention, the composition obtained by the method of the fifth aspect of the invention, the pharmaceutical composition of the sixth aspect of the invention, or the food or nutritional supplement of the seventh aspect of the invention, for use in medicine.

In a ninth aspect, the invention relates to the composition or kit of parts of the fourth aspect of the invention, the composition obtained by the method of the fifth aspect of the invention, the pharmaceutical composition of the sixth aspect of the invention, or the food or nutritional supplement of the seventh aspect of the invention, for use in the treatment of a cardiovascular disease.

BRIEF DESCRIPTION OF THE DRAWINGS

Figure 1. Diagram summarizing the study.

Figure 2. Ratio of oxLDL-cholesterol/LDL-cholesterol of the subjects who took the placebo or the extract at the start, after 4 weeks, and after 8 weeks of treatment. The mean values are represented with dots (dark gray for the control group and light gray for the extract group), and the standard error is represented by means of vertical bars. The different letters indicate significant differences between the times of each group. The asterisk (*) indicates p <0.05 between groups at the corresponding time. Figure 3. A- Chromatograms at 280 nm resolving olive tree extract polyphenols (bottom chromatogram) and almond extract polyphenols (top chromatogram). B- Chromatograms at 330 nm resolving olive tree extract polyphenols (top chromatogram) and almond extract polyphenols (bottom chromatogram).

Figure 4. A- Chromatograms at 280 nm resolving olive tree extract polyphenols (top chromatogram) and grapefruit extract polyphenols (bottom chromatogram). B-

Chromatograms at 330 nm resolving olive tree extract polyphenols (top chromatogram) and grapefruit extract polyphenols (bottom chromatogram).

Figure 5. A- Chromatograms at 280 nm resolving olive tree extract polyphenols (top chromatogram) and flax extract polyphenols (bottom chromatogram). B- Chromatograms at 330 nm resolving olive tree extract polyphenols (top chromatogram) and flax extract polyphenols (bottom chromatogram).

Figure 6. A- Chromatograms at 280 nm resolving olive tree extract polyphenols (top chromatogram) and polygonum extract polyphenols (bottom chromatogram). B-

Chromatograms at 330 nm resolving olive tree extract polyphenols (top chromatogram) and polygonum extract polyphenols (bottom chromatogram).

DETAILED DESCRIPTION OF THE INVENTION

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

Use of a polyphenol other than hydroxytyrosol for use in increasing the bioavailability and the antiatherosclerotic activity of hydroxytyrosol, and hydroxytyrosol for use in the treatment of atherosclerosis by means of its administration with a polyphenol that is not hydroxytyrosol In a first aspect, the invention relates to the use of at least one polyphenol that is not hydroxytyrosol and/or that does not occur naturally in the tree Olea europaea ( O . europaea ) for increasing the bioavailability of hydroxytyrosol after the oral administration of hydroxytyrosol or for increasing the antiatherosclerotic effect of hydroxytyrosol after its oral administration. Alternatively, the first aspect of the invention relates to a method for increasing the bioavailability of hydroxytyrosol after its oral administration or for increasing the antiatherosclerotic effect of hydroxytyrosol after its oral administration comprising the administration of at least one polyphenol that is not hydroxytyrosol and/or that does not occur naturally in the tree Olea europaea (O. europaea).

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

As it is used in the present invention, the term “hydroxytyrosol” refers to the polyphenolic compound commonly known to one skilled in the art with said name. In nature, hydroxytyrosol is present in the olive tree (e.g. in the species Olea europaea ) concentrated primarily in the leaves and in the fruit. In a particular embodiment, it refers to the chemical compound with IUPAC name 4-(2-hydroxyethyl)benzene-l,2-diol.

As it is used in the present invention, the expression “polyphenol that is not hydroxytyrosol and/or that does not occur naturally in the tree Olea europaea”, or “polyphenol that is not hydroxytyrosol and/or wherein the at least one polyphenol other than hydroxytyrosol does not occur naturally in the tree O. europaea” refers to a polyphenol, as defined above, other than hydroxytyrosol (i.e., the characteristics of which do not coincide with those of the product described in the definition of hydroxytyrosol above). Alternatively or additionally, it refers to a polyphenol which is not synthesized by the tree of the species O. europaea naturally (i.e., without the need for man to intervene in generating and selecting the tree belonging to said species or in the cultivation thereof), regardless of the cultivation method used, preferably regardless of the variety of O. europaea , more preferably regardless of the individual of said spice from which said extract is obtained. Therefore, in a particular embodiment, the polyphenol not occurring naturally in the tree O. europaea is that which is not synthesized by the tree of the species O. europaea as was just described. In a particular embodiment, the tree of the species O. europaea is the common olive tree wild variety, i.e., the variety Olea europaea sylvestris. In another particular embodiment, the tree of the species O. europaea is from 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 species O. europaea is any variety of said subspecies. In another particular embodiment, the tree is any organism of one of the taxonomic ranks of O. europaea mentioned. In a particular embodiment, the species of O. europaea mentioned in any aspect of this invention refers to the species O. europaea L.

In a particular embodiment, the polyphenol that is not hydroxytyrosol and that optionally does not occur naturally either in the tree Olea europaea is referred to in any aspect of the present invention with the expression “polyphenol that is not hydroxytyrosol”, “polyphenol other than hydroxytyrosol”, or “polyphenol differing from hydroxytyrosol”.

Methods for determining if a polyphenol occurs naturally in a tree of the species O. europaea , or is synthesized by said tree naturally, as defined above, include obtaining plant extracts from at least one plant product of a tree as described in said definition, and determining the presence of polyphenols in said extract. The absence of said polyphenol in the extracts is indicative that it is not found in said tree nor is it synthesized by said tree. Methods for obtaining plant extracts from a plant product are specified below. Methods for determining the polyphenols present in a plant extract are specified below. The definitions of plant extract and of plant product are also provided below. In a particular embodiment, the polyphenol that is not hydroxytyrosol and/or that does not occur naturally in the tree O. europaea, does not occur naturally in the fruits or in the leaves of the tree O. europaea, preferably it does not occur naturally in the leaves of the tree O. europaea. As one skilled in the art will understand, methods for determining if a polyphenol occurs naturally in the fruits and/or in the leaves of a tree O. europaea include the previously indicated methods, in which the plant extracts were obtained from the corresponding plant product, i.e., fruits and/or leaves of the tree O. europaea.

As it is used in the present invention, the expression “polyphenol” refers to the group of chemical substances commonly known to one skilled in the art by said name. It specifically refers to the chemical substances found in plants characterized by the presence of one or several phenol rings and conjugated double bonds. They are found mainly in conjugated forms, associated with carbohydrates bound to -OH groups, although there are also direct bonds of the carbohydrate with the phenol group. It is also common for them to be associated with other compounds, such as carboxylic and organic acids, amines, lipids, and even with other phenol groups. Polyphenols suitable for use in the present invention include, without limitation, phenolic acids (hydroxybenzoic acid or hydroxycinnamic acid derivatives), stilbenes, lignans, phenolic alcohols, and flavonoids, the latter constituting the most abundant subclass. Said groups of polyphenols are also used in the present invention for referring to the groups of polyphenols commonly known to one skilled in the art by the corresponding name.

Specifically, as it is used in the present invention the term “flavonoid” refers to the most abundant subclass of polyphenols in the plant kingdom. The chemical structure of flavonoids consists of a diphenylpropane backbone (C6-C3-C6) formed by two aromatic rings bound through three carbon atoms forming an oxygenated heterocycle (C). There are several subgroups of flavonoids and the classification of these compounds is done depending on the state of oxidation of the heterocyclic ring (ring C) and on the position of ring B. In each family there is a wide range of compounds differing from one another by the number and the position of the OH groups, and by the different functional groups they may present (methyls, 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 foods are flavan-3-ols, and these can occur as anthocyanidin monomers (catechin, epicatechin, gallocatechin, and epigallocatechin), as dimers condensed to one another, or oligomers (procyanidins), or they can occur as polymers (proanthocyanidins or condensed tannins). Said compounds and groups of compounds are those commonly known to one skilled in the art by the corresponding name. Non-limiting examples of flavonoids include: procyanidins, propelargonidins, prodelphinidins, catechin, epicatechin, quercetin, kaempferol, isorhamnetin, rutin, and geni stein.

As it is used in the present invention, the term “lignan” refers to phenolic compound dimers, phenylalanine derivatives, and cinnamic alcohols the basic structure of which consists of two C6C3 (n-propylbenzene) units attached by b-b’ bonds.

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. Said groups of compounds are those commonly known to one skilled in the art by the corresponding name. Non-limiting examples of lignans include: secoisolariciresinol diglucoside, secoisolariciresinol, matairesinoside, lariciresinol, pinoresinol, sesamin, schisandrin, deoxyschisandrin, gomisins, pregomisin, sesamolin, sesamol, sesaminol, sesamolinol, or pinoresinol.

As it is used in the present invention, the term “stilbene” refers to polyphenolic compounds of formula C14H12, for which there are two isomers: Irans- 1 ,2-diphenylethylene (E-stilbene) and 67 s- 1 ,2-diphenylethylene (Z-stilbene). The term stilbene includes hydroxy and alkoxy derivatives of simple stilbene, as well as its heterosides (glycosides) forms and polymers. In a particular embodiment, it refers to the chemical compound with IUPAC name: trans-1,2- diphenylethylene. Non-limiting examples of stilbenes include resveratrol, piceatannol, pinosylvin, pterostilbene, rhapontienin, pterostilbene, isorhapontienin, rhapontin, ponticin, piceid, or astringin.

As it is used in the present invention, the term “phenolic acid” or “phenolcarboxylic acid” refers to phenolic compounds containing a phenol ring and an organic function of carboxylic acid (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. As it is used in the present invention, the term “phenolic alcohol” refers to phenolic compounds comprising the alcohol function and the phenol function. Non-limiting examples of phenolic alcohols include coniferyl alcohol, sinpyl alcohol, or p-coumaryl alcohol.

In a preferred embodiment, the hydroxytyrosol for use according to the invention or the polyphenols used in the present invention are found as part of a plant extract.

As it is used in the present invention, the expression “plant extract” or “extract” refers to the term commonly known by one skilled in the art. It refers to a composition comprising compounds, ingredients, or substances, which have been extracted from a product, or tissue of a plant organism, a tree, or a plant, by means of methods commonly known to one skilled in the art. Non-limiting examples of said methods include the expression in which the plant product is introduced in a hydraulic press and squeezed, the distillation or the incision in the plant product of interest followed by the extraction of a sample of the content or composition of said plant product. Another common method for obtaining plant extracts consists of treating the product or tissue from which the plant extract is to be obtained with solvents. Non-limiting examples of solvents include water, ethanol, hydroalcohol, ethyl acetate, CO2, methanol, acetone, acetic acid, or hexane.

In a particular embodiment, the plant extract is obtained from fresh or dry plant product or tissue of interest with a solvent at a temperature of about 40 to 100°C for 1-10 hours. In a particular embodiment, the solvent is aqueous, preferably water. In another particular embodiment, the solvent is a hydroalcohol preferably having an alcohol content between 10 and 96%, wherein the alcohol is preferably ethanol or methanol. The process can also include several extraction phases for extracting the plant product with the same or different solvent, with the same or different alcohol content, combining the extracts obtained to continue the process. In another particular embodiment, the process also includes several concentration steps by means of chromatography using polymeric adsorbent resins. The obtained extract is then treated with activated carbon and then removed. Finally, and alternatively, a drying step for drying the purified extract is performed.

Therefore, in a particular embodiment, the method for obtaining the plant extract comprises: (i) extracting the fresh or dry plant product or tissue of interest with a solvent at a temperature of about 40 to 100°C for 1-10 hours (ii) treating the extract obtained in step (i) with activated carbon and then removing the activated carbon.

In another particular embodiment, the method for obtaining the plant extract comprises:

(i) extracting the fresh or dry plant product or tissue of interest with a solvent at a temperature of about 40 to 100°C for 1-10 hours

(ii) concentrating same by means of chromatography using polymeric adsorbent resins

(iii) treating the extract obtained in step (i) with activated carbon and then removing the activated carbon.

In a particular embodiment, step (i) of any of the preceding methods includes several extraction phases for extracting the plant product with the initial solvent or a different solvent, combining the extracts obtained to continue the process. If the solvent is a hydroalcohol, the alcohol content in these extraction phases is equal to or different from the initial solvent. In another particular embodiment, any of the previously described methods include a drying step for drying the extract.

In a particular embodiment, the plant extract of one or several products of O. europaea described in any aspect of the present invention, is obtained by means of the method for obtaining plant extracts with solvents described in any of the preceding embodiments, wherein the solvent is aqueous, preferably water.

In another particular embodiment, the plant extract of one or several products oiPrunus dulcis described in any aspect of the present invention, is obtained by means of the method for obtaining plant extracts with solvents described in any of the preceding embodiments, wherein the solvent is aqueous, preferably water.

In another particular embodiment, the plant extract of one or several products of Citrus paradisi described in any aspect of the present invention, is obtained by means of the method for obtaining plant extracts with solvents described in any of the preceding embodiments, wherein the solvent is a hydroalcohol.

In another particular embodiment, the plant extract of one or several products of Linum usitatissimum described in any aspect of the present invention, is obtained by means of the method for obtaining plant extracts with solvents described in any of the preceding embodiments, wherein the solvent is a hydroalcohol.

In another particular embodiment, the plant extract of one or several products of Polygonum cuspidatum described in any aspect of the present invention, is obtained by means of the method for obtaining plant extracts with solvents described in any of the preceding embodiments, wherein the solvent is a hydroalcohol.

As it is used herein, the term “activated charcoal”, “activated carbon” refers to the term well known to an expert in the field. It is a form of carbon that is processed to have augmented adsorption properties. It is mostly processed to have small pores, of low volume which increases the surface area available for adsorption or chemical reaction.

As it is used in the present invention, the term “product of a plant organism” or “plant product” refers to any part of a plant organism, including a tissue of the plant organism, a tissue of the reproductive organs of the plant organism, a tissue of the non-reproductive organs of the plant organism, the leaves, the stem, the roots, the fruits, a tissue of the fruits, the exocarp of the fruit, the mesocarp of the fruit, the endocarp of the fruit, the skin of the fruit, the shell of the fruit, the pod of the fruit, the seed of the fruit, or the pod of the plant organism. In a particular embodiment, the plant product refers to the total of any of the parts of a plant organism indicated just above. In another particular embodiment, it refers to a portion of any of the parts of the plant organism indicated just above.

Methods for determining the hydroxytyrosol content in the plant extract or the polyphenol content in the plant extract are commonly known to one skilled in the art. Said methods include any method that allows determining the amount and type of a compound in a composition, such as mass spectrometry methods. Preferably used is mass spectrometry, high-performance liquid chromatography coupled to mass spectrometry (HPLC-MS), high-performance liquid chromatography (HPLC) coupled to an ultraviolet absorbance detector, gas chromatography coupled to mass spectrometry (GC-MS), liquid chromatography coupled to mass spectrometry (LC-MS), direct infusion mass spectrometry or Fourier-transform ion cyclotron resonance mass spectrometry (FT-ICR-MS), capillary electrophoresis coupled to mass spectrometry (CE-MS), ultra-high performance liquid chromatography coupled to mass spectrometry (UHPLC-MS), supercritical fluid chromatography coupled to mass spectrometry (SFC-MS), flow injection analysis with mass spectrometry (FIA-MS), including quadrupole mass spectrometry, any sequentially coupled 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 zero. Said techniques are disclosed in, for example, Nissen, Journal of Chromatography A, 703, 1995: 37-57, documents US 4,540,884 or US 5,397,894. Preferably, gas chromatography coupled to mass spectrometry as described in (Garcia A. and Barbas C. 2011, Methods Mol. Biol. (Clifton NJ) 708:191-204) is used. Even more preferably, quadrupole time-of-flight gas chromatography coupled to mass spectrometry (GC-qTOF/MS) as described in (Riera-Borrull M. etal. 2016, J. Am. Soc. Mass Spectrom. 27(1): 168-177; Kind T. et al, 2009, Anal. Chem. 81(24): 10038-48) is used. In a preferred embodiment, the determination of the levels of hydroxytyrosol in a sample is carried out by means of any of the aforementioned chromatographic techniques using a calibration line obtained by means of a hydroxytyrosol pattern of a known concentration.

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

As it is used in the present invention, the expression “bioavailability” refers to the fraction and the rate at which the administered dose of a compound reaches its therapeutic target (cell receptors, channels, carriers), which implies reaching the tissue on which it acts. It is considered equivalent to the levels reached in the systemic circulation of a subject who has ingested or who has been administered said compound. This is why, in practice, bioavailability is determined by the percentage of compound occurring in plasma after its administration. As it is used in the present invention, the expression “bioavailability of hydroxytyrosol” refers to the bioavailability as defined above, wherein the compound is hydroxytyrosol. Therefore, the expression “bioavailability of hydroxytyrosol after its oral administration” refers to the percentage of hydroxytyrosol in systemic circulation, or in plasma, once hydroxytyrosol has been administered orally to a subject. As one skilled in the art will understand, the more stable hydroxytyrosol is in the digestive tract, the higher the amount of hydroxytyrosol is available in the intestinal lumen for absorption, and the greater the amount of hydroxytyrosol that reaches the blood supply. Therefore, the greater the stability of the hydroxytyrosol is in the digestive tract after its oral administration, the greater its bioavailability is. Therefore, methods which allow determining the bioavailability of hydroxytyrosol after its oral administration comprise determining the percentage of hydroxytyrosol in serum of a subject who has been administered hydroxytyrosol, or determining the stability of the hydroxytyrosol in the digestive tract of a subject who has been administered hydroxytyrosol orally. Non limiting examples of methods for determining the percentage of hydroxytyrosol in serum of a subject include any of the methods indicated above for determining the presence of polyphenols in a plant extract, applied to a serum sample from said subject. Non-limiting examples of methods which allow determining the stability of hydroxytyrosol in the digestive tract include the method described in the section B on materials and methods and in Example 3 of the application. Said methods can also be variants of those described in said sections of the application, specifically, the step for detecting the content of hydroxytyrosol in each of the compositions of the method of the application may consist of any of the methods indicated above for detecting polyphenols in a plant extract (wherein the polyphenol is hydroxytyrosol, and the plant extract is substituted with the corresponding composition of the method of the application in which hydroxytyrosol has been incubated).

As it is used in the present invention, the expression “increasing the bioavailability of hydroxytyrosol after its oral administration” refers to increasing 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% the bioavailability of hydroxytyrosol in a subject with respect to a reference value. In a particular embodiment, said reference value is the bioavailability of hydroxytyrosol after the oral administration of hydroxytyrosol without a polyphenol that is not hydroxytyrosol and that does not occur naturally in the tree O. europaea,, as said polyphenol is defined and described above and in the different embodiments of the present invention. As one skilled in the art will understand, said reference value is the bioavailability of hydroxytyrosol after the oral administration of a compound the active ingredient of which is only hydroxytyrosol. Said compound may consist of a composition consisting of hydroxytyrosol and at least one pharmaceutically acceptable excipient, wherein said excipient can be any of those indicated in the aspect of the invention relating to the pharmaceutical composition of the invention. Methods for determining the bioavailability of hydroxytyrosol, such as those which have been described just above, allow determining if the bioavailability of hydroxytyrosol is higher or lower, and at what percentage, with respect to a reference value. As one skilled in the art will understand, said methods also allow measuring the bioavailability of hydroxytyrosol defined as a reference value and therefore determining the reference value indicated in the present definition.

The term “atherosclerosis”, “atherosclerotic vascular disease”, or “ASVD”, refers to the cardiovascular disease characterized by chronic inflammation, the accumulation of lipids, modifications of vascular cells in the arterial wall, and the formation of atherosclerotic plaque or atheroma. Said processes trigger a narrowing of the arterial lumen. Oxidized low-density lipoproteins (oxLDL) are considered to play a key role in the initiation and progression of atherogenesis. oxLDLs bind with a higher affinity than native LDL to several receptors, so their absorption by macrophages is more rapid, which leads to the buildup of cholesterol and the formation of atherosclerotic plaque, or atheroma. Furthermore, oxLDL promotes endothelial cell activation and dysfunction, vascular smooth muscle cell migration and proliferation, and platelet activation, causing an inflammatory response, endothelial dysfunction; aggravating the atherogenic process. To that end, the concentration of circulating oxLDL is considered one of the most important biomarkers of the progression of atherosclerosis.

As it is used in the present invention, the expression “antiatherosclerotic effect of hydroxytyrosol” refers to the activity of hydroxytyrosol once administered to a subject, which helps to prevent, cur, reverse, or treat atherosclerosis. Suitable indicators of the antiatherosclerotic effect of hydroxytyrosol include the reduction in the concentration of serum oxLDL, or the reduction of the serum oxLDL/LDL-cholesterol ratio, once administered to a subject, preferably orally. As it is used in the present invention, the expression “LDL”, “LDL-c” or “LDL-cholesterol” refers to the term commonly known by one skilled in the art; it specifically refers to low- density lipoproteins or LDLs. LDLs are one of the 5 main groups of lipoproteins which transport fat molecules in the body in the extracellular medium (ULDs, ultra-low-density lipoproteins; VLDL, very low-density lipoprotein; IDL intermediate-density lipoprotein; HDL, high-density lipoprotein; and LDL as it is here defined). Most of the cholesterol is transported in the blood of a subject along with proteins, forming LDLs.

As it is used in the present invention, the expression “ox-LDL” or “oxLDL” refers to LDL lipoproteins as defined above, which are oxidized. Oxidized LDL is a general term for LDL lipoproteins with structural components modified by oxidation. As a result of the attack of free radicals, both the lipid parts and the protein parts of LDL can be oxidized. In addition to the oxidative reactions taking place in the vascular wall giving rise to ox-LDL, lipids oxidized into LDL can also derive from dietary oxidized lipids. ox-LDL is associated with the development of atherosclerosis. The effect of oxidized LDLs on atherogenesis has been explained by the lack of recognition by LDL receptors of the structures of LDL modified by oxidation, which prevents the development of the normal metabolism of the LDL particles and finally leads to the development of atherosclerotic plaques.

As one skilled in the art will understand that as it is used in the present invention, the expression “ox-LDL/LDL-cholesterol” or “ox-LDL/LDL” refers to the ratio between the concentration in a specific ox-LDL medium as defined above and the concentration of LDL as defined above in said medium.

As it is used in the present invention, the expression “increasing the antiatherosclerotic effect of hydroxytyrosol” refers to increasing 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% any of the effects indicated in the particular embodiments above in the definition of atherosclerotic effect with respect to a reference value. In a particular embodiment, the reference value is the value of the parameter changed by the administration of hydroxytyrosol indicated in the corresponding preceding embodiment in the definition of atherosclerotic effect, upon administration of hydroxytyrosol without a polyphenol that is not hydroxy tyro sol and that does not occur naturally in the tree O. europaea, as said polyphenol is defined and described above and in the different embodiments of the present invention. Preferably, the hydroxytyrosol administered for determining the reference value consists of a composition the active compound of which is only hydroxytyrosol, specifically comprising only hydroxytyrosol and optionally a pharmaceutically acceptable excipient, as defined in the aspect of the invention relating to the pharmaceutical composition of the invention.

In a particular embodiment, the expression “increasing the antiatherosclerotic effect of hydroxytyrosol” refers to decreasing 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 by at least 20% the serum oxLDL concentration with respect to a reference value. In a particular embodiment, said decrease with respect to the reference value is of at least 50%. In another particular embodiment, it is of at least 70%. In a particular embodiment, said reference value is the serum oxLDL concentration after the oral administration of hydroxytyrosol without a polyphenol that is not hydroxytyrosol and that does not occur naturally in the tree O. europaea, as said polyphenol is defined and described above and in the different embodiments of the present invention. Preferably, the hydroxytyrosol administered for determining the reference value consists of a composition the active compound of which is only hydroxytyrosol, specifically comprising only hydroxytyrosol and optionally a pharmaceutically acceptable excipient, as defined in the aspect of the invention relating to the pharmaceutical composition of the invention. Methods for determining said decrease include the methods described in section A- on materials and methods (particularly in the “Analysis of biochemical parameters” section) of the application for determining the serum oxLDL concentration and comparing the value obtained with the reference value. In another particular embodiment, the expression “increasing the antiatherosclerotic effect of hydroxytyrosol” refers to decreasing 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 by at least 25% the serum oxLDL/LDL- cholesterol ratio with respect to a reference value. In a particular embodiment, said decrease with respect to the reference value is of at least 30%. In another particular embodiment, it is of at least 50%. In a particular embodiment, said reference value is the serum oxLD/LDL- cholesterol ratio after the oral administration of hydroxytyrosol without a polyphenol that is not hydroxytyrosol and that does not occur naturally in the tree O. europaea, as said polyphenol is defined and described above and in the different embodiments of the present invention. Preferably, the hydroxytyrosol administered for determining the reference value consists of a composition the active compound of which is only hydroxytyrosol, specifically comprising only hydroxytyrosol and optionally a pharmaceutically acceptable excipient, as defined in the aspect of the invention relating to the pharmaceutical composition of the invention. Methods for determining said decrease include the methods for determining the serum oxLDL concentration described in section A- on materials and methods (particularly in the “Analysis of biochemical parameters” section) of the application, and methods for determining the serum LDL concentration commonly known to one person skilled in the art. Non-limiting examples of said methods include the direct method (G.D. 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 (Wamick GR et al. Lab Med. 2008;39(8):481-90). Then the ratio between said values is determined, and the value or values of the oxLDL/LDL-cholesterol ratio obtained in a sample of serum of interest are compared with the reference value.

In particular 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 occur naturally in Olea europaea is administered in combination with hydroxytyrosol, i.e., as part of one and the same composition. Alternatively, in particular embodiments of the first, second, and third aspects of the invention, the at least one polyphenol that is not hydroxytyrosol as such and/or that does not occur naturally in Olea europaea is administered sequentially with respect to hydroxytyrosol. Alternatively, in particular embodiments of the first, second, and third aspects of the invention, the at least one polyphenol that is not hydroxytyrosol as such and/or that does not occur naturally in Olea europaea is administered separately with respect to the hydroxytyrosol. In a particular embodiment of the first, second, and third aspects of the invention, the hydroxytyrosol is part of a plant extract.

The expression “plant extract” has been defined above. In a particular embodiment, the plant extract comprising hydroxytyrosol is obtained from any plant product of the tree O. europaea, wherein the plant product is any of those indicated in the definition of “plant product” indicated above. In a preferred embodiment of the first, second, and third aspects of the invention, the plant extract of which the hydroxytyrosol is part is an extract of the leaf or of the fruit, preferably the leaf, of the tree O. europaea.

As it is used in the present invention, the expression “is part of a plant extract” refers to a specific compound being comprised in a plant extract, wherein the term “comprised”, “comprises”, indicates that the plant extract must contain said compound, but it may optionally contain additional compounds or ingredients.

In a particular embodiment, the plant extract comprising hydroxytyrosol has been obtained from at least two plant products of the tree O. europaea , selected from those indicated in the definition of plant product, wherein each of said products is the same or different. In another particular embodiment, each of said products is obtained from a different tree of O. europaea , wherein each tree is preferably of a different subspecies, more preferably wherein each tree is of a different variety.

In a particular embodiment, the tree of the species O. europaea mentioned in any definition or particular embodiment of any aspect of the present invention, is any variety of the tree of the spice O. europaea. In another particular embodiment, it is the common olive tree wild variety, i.e., the variety Olea Europaea sylvestris. In another embodiment, it is a tree 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, it is any variety of said subspecies.

In a particular embodiment, the trees of the species O. europaea mentioned in any definition or particular embodiment of any aspect of the present invention, are each of any variety of the tree of the spice O. europaea. In another particular embodiment, at least one is the common olive tree wild variety, i.e., the variety Olea Europaea sylvestris. In another embodiment, at least one is a tree 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 at least one is of any variety of said subspecies.

In a particular embodiment, the plant extract comprising hydroxytyrosol contains 1-100% (w/w), 1-98% (w/w), 1-95% (w/w), 1-90% (w/w), 3-90% (w/w), 5-90% (w/w), 7-90% (w/w), 8-90% (w/w), 9-90% (w/w), 10-90% (w/w), 10-85% (w/w), 10-80%(w/w), 10-70% (w/w), 10-60% (w/w), 10-50% (w/w), 10-40% (w/w), 10-30% (w/w), 10-25% (w/w), 10-20% (w/w) of hydroxytyrosol. In a preferred embodiment, the plant extract comprising hydroxytyrosol contains 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), 99.9% (w/w), 99.95% (w/w), 9.99% (w/w), 100% (w/w), preferably 10% (w/w) of hydroxytyrosol.

In a particular embodiment of the first, second, and third aspects of the invention, the plant extract comprising hydroxytyrosol contains 10-90%, preferably 10% (w/w) of hydroxytyrosol.

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.

As used in the present invention, the terms “flavonoids”, “lignans” and “stilbenes” refer to the polyphenols commonly known by said name to one skilled in the art. Specifically, they refer to polyphenols as defined above.

In a particular embodiment of the first, second, and third aspects of the invention, the at least one polyphenol other than hydroxytyrosol of the group of flavonoids is selected from the group consisting of procyanidins, propelargonidins, prodelphinidins, catechin, epicatechin, quercetin, kaempferol, isorhamnetin, naringin, naringenin, genistein, and rutin. As used in the present invention, the terms “procyanidins”, “propelargonidins”, “prodelphinidins”, “catechin”, “epicatechin”, “quercetin”, “kaempferol”, “isorhamnetin”, “naringin”, “naringenin”, “genistein”, and “rutin” refer to the corresponding term as understood by one skilled in the art.

As it is used in the present invention, the term “procyanidins” refers to the group of proanthocyanidins constituted exclusively of (epi)catechin. As it is used in the present invention, the term “propelargonidins” refers to proanthocyanidins containing an (epi)afzelechin unit together with (epi)catechin units. As it is used in the present invention, the term “prodelphinidins” refers to proanthocyanidins containing at least one (epi)gallocatechin unit together with (epi)catechin units. As it is used in the present invention, the term “proanthocyanidin” or “condensed tannins” refers to flavan-3-ol oligomers and polymers widely distributed in the plant kingdom. The most abundant flavan-3-ol units are (+)-afzelechin, (+)-catechin, and (+)-gallocatechin (2R:3S forms) and their diastereoisomers (-)-epiafzelechin, (-)epi catechin, and (-)-epigallocatechin (2R:3R forms), respectively. In type B procyanidins/propelargonidins/prodelphinidins, the flavan-3-ol units are bound by C-4 carbon of the upper unit and C-6 or C-8 carbon of the lower unit, C4-C8 isomers being more abundant than C4-C6 isomers. In addition to this interflavan C-C bond, type A 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 L.J. (1988) Flavans and proanthocyanidins. In The flavonoids (Harbome J.B., Ed.) Chapman and Hall, New York, pp. 21-62).

Therefore, in a particular embodiment, the polyphenol other than hydroxytyrosol of the group of flavonoids referred to in any aspect of the present invention, can be any polyphenol belonging to the group of proanthocyanidins. In a particular embodiment, it is any polyphenol of the group of procyanidins, preferably it is a type B procyanidin. In a particular embodiment, it is a procyanidin selected from the group consisting of type B1 procyanidin, type B2 procyanidin, type B3 procyanidin, type B4 procyanidin, type B5 procyanidin, type B6 procyanidin, type B7 procyanidin. In another particular embodiment, it is a procyanidin of type Cl. In a particular embodiment, it is a procyanidin of type A, preferably selected from the group consisting of procyanidin Al, procyanidin A2. In another particular embodiment, it is any polyphenol of the group of propelargonidins, preferably type A propelargonidins. In another particular embodiment, it is a type A propelargonidin, more preferably it is a type B propelargonidin. In another particular embodiment, it is any polyphenol of the group of prodelphinidins; in a particular embodiment, preferably type A prodelphinidins.

Specifically, as it is used in the present invention, the term “procyanidin Bl” or “type B1 procyanidin” refers to the compound with IUPAC name: (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.

As it is used in the present invention, the term “procyanidin B2” or “type B2 procyanidin” refers to the compound with 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.

As it is used in the present invention, the term “procyanidin B3” or “type B3 procyanidin” refers to the compound with 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.

As it is used in the present invention, the term “procyanidin B4” or “type B3 procyanidin” refers to the compound with 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.

As it is used in the present invention, the term “procyanidin B5” or “type B5 procyanidin” refers to the compound with 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.

As it is used in the present invention, the term “procyanidin B6” or “type B6 procyanidin” refers to the compound with 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. The term “procyanidin B7”, or “procyanidin type B7”, as it is used in the present invention, refers to the compound with name (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- chromene-3,5,7-triol.

As it is used in the present invention, the term “procyanidin Cl” or “type Cl procyanidin” refers to the compound with 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.

As it is used in the present invention, the term “procyanidin Al” or “type A1 procyanidin” refers to the compound with IUPAC name: (lR,5R,6S,13S,21R)-5,13-bis(3,4- dihydroxyphenyl)-4, 12, 14-trioxapentacyclo[l 1.7.1.02, 11.03,8.015, 20]henicosa- 2( 11 ),3(8),9, 15, 17, 19-hexaene-6,9, 17,19,21 -pentol .

As it is used in the present invention, the term “procyanidin A2” or “type A2 procyanidin” refers to the compound with IUPAC name (lR,5R,6R,13S,21R)-5,13-bis(3,4- dihydroxyphenyl)-4, 12, 14-trioxapentacyclo[l 1.7.1.02, 11.03,8.015, 20]henicosa- 2( 11 ),3(8),9, 15, 17, 19-hexaene-6,9, 17,19,21 -pentol .

As it is used in the present invention, the term “propelargonidin” refers to the polyphenol with 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-tetrol.

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

As used in the present invention, the terms “secoisolariciresinol diglucoside”, “matairesinoside”, “lariciresinol”, “pinoresinol”, “sesamin”, and “secoisolariciresinol” refer to the corresponding term as understood by one skilled in the art. In another particular embodiment of the first, second, and third aspects of the invention, the at least one polyphenol other than hydroxytyrosol of the group of stilbenoids is selected from the group consisting of resveratrol, piceatannol, pinosylvin, pterostilbene, and rhapontienin.

As used in the present invention, the terms “resveratrol”, “piceatannol”, “pinosylvin”, “pterostilbene”, and “rhapontienin” refer to the corresponding term as understood by one skilled in the art.

In a particular embodiment of the first, second, and third aspects of the invention, the at least one polyphenol other than hydroxytyrosol is part of a plant extract.

In another particular embodiment, said plant extract is not an extract of a plant product of the tree O. europaea , preferably it is not an extract of the leaf or of the fruit of the tree (). europaea.

In another particular embodiment of the first, second, and third aspects of the invention, the plant extract of which the at least one polyphenol other than hydroxytyrosol is part is an extract selected from the group consisting of an extract of a plant product of Prunus dulcis (Miller) D.A. Webb an extract of a plant product of Citrus paradisi MacFad an extract of a plant product of Linum usitatissimum L an extract of a plant product of Polygonum cuspidatum Sieb. et Zucc

In a particular embodiment, each of the plant products mentioned in the preceding embodiment is one of those indicated in the definition of “plant product” provided above.

In a preferred embodiment, the plant extract of which the at least one polyphenol other than hydroxytyrosol is part is an extract selected from the group consisting of an extract of the skin of the fruit of Prunus dulcis (Miller) D.A. Webb an extract of the fruit of Citrus paradisi MacFad or an extract of the seed of the fruit of Citrus paradisi MacFad an extract of the seed of the fruit of Linum usitatissimum L an extract of the root of Polygonum cuspidatum Sieb. et Zucc. The plant extract of which the at least one polyphenol other than hydroxytyrosol is part specified in the preceding embodiments is obtained from any plant or vegetable belonging to the species indicated in each case, preferably of any variety belonging to the species indicated in each case. In a particular embodiment, the tree is of the species Prunus dulcis (Miller) D. A. Webb.

In a particular embodiment, the term Prunus dulcis used in any aspect of the invention is interchangeable with the term Prunus dulcis (Miller) D.A. Webb. In another particular embodiment, the term Citrus paradisi used in any aspect of the invention is interchangeable with the term Citrus paradisi MacFad. In another particular embodiment, the term Linum usitatissimum used in any aspect of the invention is interchangeable with the term Linum usitatissimum L. In another particular embodiment, the term Polygonum cuspidatum used in any aspect of the invention is interchangeable with the term Polygonum cuspidatum Sieb. et Zucc.

In a particular embodiment, the plant extract comprising the polyphenol other than hydroxytyrosol obtained from a specific species, is obtained from more than one plant product of the tree, plant or vegetable of said species, wherein each of said products is the same or different. In another particular embodiment, each of said products is obtained from different trees, plants, or vegetables, wherein each of said trees, plants, or vegetables are of the same species, preferably of the same variety.

Therefore, in a particular embodiment, the plant extract of which the at least one polyphenol other than hydroxytyrosol is part is an extract selected from the group consisting of an extract of at least two plant products, each selected from the list of plant products indicated in the definition of “plant product” above and obtained from at least two different trees of Prunus dulcis, wherein each tree is preferably of a different variety an extract of at least two plant products, each selected from the list of plant products indicated in the definition of “plant product” above and obtained from at least two different trees of Citrus paradisi, wherein each tree is preferably of a different variety an extract of at least two plant products, each selected from the list of plant products indicated in the definition of “plant product” above and obtained from at least two different plants of Linum usitatissimum, wherein each plant is preferably of a different variety. an extract of at least two plant products, each selected from the list of plant products indicated in the definition of “plant product” above and obtained from at least two different plants of Polygonum cuspidatum, wherein each plant is preferably of a different variety.

In particular embodiments: the at least one polyphenol other than hydroxytyrosol which is part of any aforementioned extract of one or several plant products, preferably the extract of the skin of the fruit, of one or several trees of Prunus is a flavonoid, preferably selected from the group consisting of procyanidins, propelargonidins, prodelphinidins, catechin, epicatechin, quercetin, kaempferol, isorhamnetin, rutin, and genistein. the at least one polyphenol other than hydroxytyrosol which is part of any aforementioned extract of one or several plant products, preferably of the seed of the fruit, of one or several trees Citrus paradisi, 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 any aforementioned extract of one or several plant products, preferably of the seed 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 which is part of any aforementioned extract of one or several plant products, preferably of the root, of one or several plants of Polygonum cuspidatum , is a stilbene, preferably selected from the group consisting of resveratrol, piceatannol, pinosylvin, pterostilbene, and rhapontienin.

In preferred embodiments of the first, second, and third aspects of the invention: the at least one polyphenol other than hydroxytyrosol which is part of the extract of the skin of the fruit of Prunus dulcis is a flavonoid, preferably selected from the group consisting of procyanidins, propelargonidins, prodelphinidins, catechin, epicatechin, quercetin, kaempferol, isorhamnetin, rutin, and genistein. the at least one polyphenol other than hydroxytyrosol which is part of the extract of the fruit of Citrus paradisi or the extract of the seed of the fruit of Citrus paradisi 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 seed of the fruit of Linum usitatissimum is a lignan, preferably selected from the group consisting of secoisolariciresinol diglucoside, secoisolariciresinol, matairesinoside, lariciresinol, pinoresinol, and sesamin.

In a particular embodiment, any aforementioned extract of one or several plant products, preferably the extract of the skin of the fruit, of one or several trees Prunus dulcis contains between 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) of flavonoids, preferably between 5-60% (w/w) of flavonoids. Preferably, said flavonoids are selected from the group consisting of procyanidins, propelargonidins, prodelphinidins, catechin, epicatechin, quercetin, kaempferol, isorhamnetin, rutin, genistein, and combinations thereof. A particular embodiment, contains 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) of flavonoids. Preferably, said flavonoids are selected from the list of flavonoids indicated previously. In a preferred embodiment, the determination of the polyphenol content is carried out using the method based on the Folin-Ciocalteu reagent (Singleton VL et al. , 1999, Methods in Enzymology , vol. 299: 152-78).

In a preferred embodiment, the extract of the skin of the fruit of Prunus dulcis contains between 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) of flavonoids, preferably between 5-60% (w/w). Preferably, said flavonoids are selected from the group consisting of procyanidins, propelargonidins, prodelphinidins, catechin, epicatechin, quercetin, kaempferol, isorhamnetin, rutin, genistein, and combinations thereof. In a particular embodiment, the extract contains 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) of flavonoids. Preferably, said flavonoids are selected from the list of flavonoids indicated previously. In a preferred embodiment, the determination of the polyphenol content is carried out using the method based on the Folin-Ciocalteu reagent (Singleton VL et al. , 1999, Methods in Enzymology , vol. 299: 152-78).

In another particular embodiment, any aforementioned extract of one or several plant products, preferably of the fruit or of the seed of the fruit, of one or several trees Citrus paradisi , contains between 1-100% (w/w), 1-90% (w/w), 5-80% (w/w), 5-70% (w/w), 5-80% (w/w), 10-80% (w/w), 10-60% (w/w), 15-50% (w/w), 20-50% (w/w), 30-50% (w/w), preferably between 10-80% (w/w) of flavonoids. Preferably, said flavonoids are selected from the list consisting of naringin, naringenin, quercetin, catechin, rutin, genistein, and combinations thereof. In a particular embodiment, the extract contains 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) of flavonoids. Preferably, said flavonoids are 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 the method based on the Folin-Ciocalteu reagent (Singleton VL et al. , 1999, Methods in enzimology , vol. 299: 152-78).

In a preferred embodiment, the extract of the fruit of Citrus paradisi or the extract of the seed of the fruit of Citrus paradisi contains between 1-100% (w/w), 1-90% (w/w), 5-80% (w/w), 5-70% (w/w), 5-80% (w/w), 10-80% (w/w), 10-60% (w/w), 15-50% (w/w), 20-50% (w/w), 30-50% (w/w), preferably between 10-80% (w/w) of flavonoids. Preferably, said flavonoids are selected from the list consisting of naringin, naringenin, quercetin, catechin, rutin, genistein, and combinations thereof. In a particular embodiment, the extract contains 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) of flavonoids. Preferably, said flavonoids are selected from the list consisting of naringin, naringenin, quercetin, catechin, rutin, geni stein, and combinations thereof. In a preferred embodiment, the determination of the polyphenol content is carried out using the method based on the Folin-Ciocalteu reagent (Singleton VL et al, 1999, Methods in Enzymology, vol. 299: 152-78).

In another particular embodiment, any aforementioned extract of one or several plant products, preferably the extract of the seed of the fruit, of one or several plants of Linum usitatissimum contains between 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) of lignans, preferably between 5-50% (w/w) lignans. Preferably, said lignans are selected from the group consisting of secoisolariciresinol diglucoside, secoisolariciresinol, matairesinoside, lariciresinol, pinoresinol, and sesamin, and combinations thereof. In a particular embodiment, the extract contains 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 20% (w/w) of lignans. Preferably, said lignans are selected from the aforementioned list. In a preferred embodiment, the determination of the polyphenol content is carried out using the method based on the Folin-Ciocalteu reagent (Singleton VL et al ., 1999, Methods in enzimology , vol. 299: 152-78).

In a preferred embodiment, the extract of the seed of Linum usitatissimum contains between 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) of lignans, preferably between 5-50% (w/w) lignans. Preferably, said lignans are selected from the group consisting of secoisolariciresinol diglucoside, secoisolariciresinol, matairesinoside, lariciresinol, pinoresinol, and sesamin, and combinations thereof. In a particular embodiment, the extract contains 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 20% (w/w) of lignans. Preferably, said lignans are selected from the aforementioned list. In a preferred embodiment, the determination of the polyphenol content is carried out using the method based on the Folin-Ciocalteu reagent (Singleton VL et al., 1999, Methods in enzimology , vol. 299: 152-78).

In another particular embodiment, any aforementioned extract of one or several plant products, preferably of the root, of one or several plants Polygonum cuspidatum, contains between 10-100% (w/w), 20-100% (w/w), 30-100% (w/w), 40-100% (w/w), 50-100% (w/w), 60-100% (w/w), 70-100% (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) of stilbenes, preferably between 50-100% (w/w) of stilbenes. Preferably, said stilbenes are selected from the list consisting of resveratrol, piceatannol, pinosylvin, pterostilbene, rhapontienin, and combinations thereof, more preferably resveratrol. In a particular embodiment, it is a combination of resveratrol and at least one of the other stilbenoids indicated in the preceding list. In a particular embodiment, 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), 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) of stilbenes. Preferably, said stilbenes are selected from the list consisting of resveratrol, piceatannol, pinosylvin, pterostilbene, rhapontienin, and combinations thereof, more preferably resveratrol. In a particular embodiment, it is a combination of resveratrol and at least one of the other stilbenoids indicated in the preceding list. In a preferred embodiment, the determination of the polyphenol content is carried out using the method based on the Folin-Ciocalteu reagent (Singleton VL et al ., 1999, Methods in Enzymology , vol. 299: 152-78).

In a preferred embodiment, the extract of the root of Polygonum cuspidatum contains between 10-100% (w/w), 20-100% (w/w), 30-100% (w/w), 40-100% (w/w), 50-100% (w/w), 60-100% (w/w), 70-100% (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) of stilbenes, preferably between 50-100% (w/w) of stilbenes. Preferably, said stilbenes are selected from the list consisting of resveratrol, piceatannol, pinosylvin, pterostilbene, rhapontienin, and combinations thereof, more preferably resveratrol. In a particular embodiment, it is a combination of resveratrol and at least one of the other stilbenoids indicated in the preceding list. In a particular embodiment, 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), 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) of stilbenes. Preferably, said stilbenes are selected from the list consisting of resveratrol, piceatannol, pinosylvin, pterostilbene, rhapontienin, and combinations thereof, more preferably resveratrol. In a particular embodiment, it is a combination of resveratrol and at least one of the other stilbenoids indicated in the preceding list. In a preferred embodiment, the determination of the polyphenol content is carried out using the method based on the Folin-Ciocalteu reagent (Singleton VL et al. , 1999, Methods in Enzymology , vol. 299: 152-78).

In preferred embodiments: the extract of the skin of the fruit of Prunus dulcis contains between 5-60% (w/w), preferably 30% (w/w) of flavonoids, wherein the flavonoids are preferably selected from the group consisting of procyanidins, propelargonidins, prodelphinidins, catechin, epicatechin, quercetin, kaempferol, isorhamnetin, rutin, genistein, and combinations thereof the extract of the fruit of Citrus paradisi or the extract of the seed of the fruit of Citrus paradisi contains between 10-80% w/w, preferably 45% (w/w) of flavonoids, wherein the flavonoids are preferably selected from the group consisting of naringin, naringenin, quercetin, catechin, rutin, genistein, and combinations thereof. the extract of the seed of the fruit of Linum usitatissimum contains between 5- 50% (w/w), preferably 20% (w/w) of lignans, wherein the lignans are preferably selected from the group consisting of secoisolariciresinol diglucoside, secoisolariciresinol, matairesinoside, lariciresinol, pinoresinol, and sesamin, and combinations thereof. the extract of the root of Polygonum cuspidatum contains between 50-100% (w/w), preferably 98% (w/w) of stilbenoids, wherein the stilbenoids are preferably selected from the group consisting of resveratrol, piceatannol, pinosylvin, pterostilbene, rhapontienin, and combinations thereof.

In a particular embodiment, the polyphenol other than hydroxytyrosol is used at least at 1.25, 1.5, 1.75, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.4, 3.5, 3.75, 4, 4.25, 4.5, 4.75, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 10, 11, 12, 51, 18, 20, 21, 22, 23, 24, 24.2, 24.3, 24.5, 25, 26, 27, 28, 29, 30, 32, 35, 40 preferably 2.7 times in excess by weight with respect to the amount by weight of the orally administered hydroxytyrosol.

In another particular embodiment, the w/w ratio of hydroxytyrosohpolyphenol other than hydroxytyrosol used in the first, second, or third aspect is between 1:0.5 and 1:100, 1:1 and 1:50, 1:2 and 1:40.1:2 and 1:30, 1:2 and 1:28, 1:2 and 1:25, 1:2 and 1:24.5, 1:2 and 1:24.3, 1:2 and 1:24, 1:2 and 1:20, 1:2 and 1: 17, 1:2 and 1:15, 1:2 and 1:10, 1: 2 and 1:7, 1:2 and 1:5, 1:2 and 1: 4, 1:2 and 1:3, preferably between 1:1 and 1:50.

In another particular embodiment, the w/w ratio of hydroxytyrosohpolyphenol other than hydroxytyrosol used in the first, second, or third aspect is of 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, even more preferably 1:28.

In a particular embodiment, the w/w ratio of plant extract comprising hydroxytyrosohplant extract comprising the at least one polyphenol other than hydroxytyrosol used in the first, second, or third aspect is between 1:0.5 and 1:100, 1:1: and 1:75, 1:2 and 1:50, 1:3 and 1:40, 1:4 and 1:30, 1:5 and 1:25, 1:7 and 1:20, 1:10 and 1: 15, preferably between 1:2 and 1:50. In another particular embodiment, the w/w ratio of plant extract comprising hydroxytyrosohplant extract comprising a polyphenol other than hydroxytyrosol is of 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.

2- Composition and kit of parts of the invention In a fourth aspect, the invention relates to a composition or kit of parts comprising hydroxytyrosol and at least one polyphenol that is not hydroxy tyro sol, at least one polyphenol that does not occur naturally in the tree O. europaea , or at least one polyphenol other than hydroxytyrosol and that does not occur naturally in the tree O. europaea.

In a particular embodiment, the hydroxytyrosol and the at least one polyphenol that is not hydroxytyrosol, the at least one polyphenol that does not occur naturally in the tree O. europaea, or at least one polyphenol other than hydroxytyrosol and that does not occur naturally in the tree O. europaea, are called the compounds of the composition of the invention.

As it is used in the present invention, the term “composition” refers to a combination of compounds or ingredients. The ingredients of the composition can be supplied separately, or in a dosage form. Therefore, if the composition is administered to a subject, the compounds of the composition could be mixed in said dosage form, mixed before administration, despite being supplied separately, being supplied and administered separately, but mixed once taken by the subject, i.e., inside the subject’s body. Furthermore, some ingredients of the composition can be administered together and others separately, but they are all mixed once taken by the subject, i.e., inside the subject’s body.

The composition according to the invention can be supplied with different formats. Non limiting examples include tablets, coated tablets, pills, aqueous or oily suspensions, solutions, dispersible powders or granules, emulsions, hard or soft capsules, syrups or elixirs, pastes, gels, or the like. Said composition can be prepared according to any known method, and such compositions may contain in addition to the compounds indicated above in the composition of the invention, one or more agents selected from the group consisting of sweetening agents, flavoring agents, coloring agents, and preserving agents to provide pharmaceutically elegant and pleasing compositions. Any of the formats in which the composition is supplied may contain the active ingredient(s) in a mixture with non-toxic pharmaceutically acceptable excipients that are suitable for manufacturing said supply formats. These excipients can be, for example, inert diluents, such as calcium carbonate, sodium carbonate, lactose, calcium phosphate, or sodium phosphate; granulating and disintegrating agents, for example, corn starch or alginic acid; binding agents, for example, starch, gelatin, or acacia gum; and lubricating agents, for example, magnesium stearate, stearic acid, or talc. The different formats in which the composition is supplied can be uncoated or coated by known techniques to delay disintegration and absorption in the digestive system and thereby provide an action sustained over a longer period. For example, a time delay material such as glyceryl monostearate or glyceryl distearate can be used. They may also be coated for controlled administration. For example, a “delayed-release” pharmaceutical form releases a product or substance at a different time rapidly after the administration. The examples of delayed-release systems include repeat-action tablets and capsules and tablets with an enteric coating where the scheduled release is achieved through a barrier coating.

The composition according to the invention may also be formulated for oral use as hard gelatin capsules, wherein the active ingredient(s) is/are mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate, or kaolin, or soft gelatin capsules, wherein the active ingredient(s) is/are mixed with water and an oily medium, for example, peanut oil, liquid paraffin, or olive oil.

The composition according to the invention can be formulated as aqueous suspensions wherein the active ingredient(s) is/are in a mixture with excipients suitable for manufacturing aqueous suspensions. Such excipients are suspension agents, for example, sodium carboxymethylcellulose, methylcellulose, hydroxypropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, tragacanth gum, and acacia gum; the dispersing or wetting agents can be a natural phosphatide such as lecithin, or products of condensation of an alkylene oxide with fatty acids, for example, polyoxyethylene stearate, or products of condensation of ethylene oxide with long-chain aliphatic alcohols, for example, heptadecaethylenoxyketanol, or products of condensation of ethylene oxide with partial esters derived from fatty acids and a hexitol such as polyoxyethylene sorbitol monooleate, or products of condensation of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, for example, polyethylene sorbitan monooleate. The aqueous suspensions may also contain one or more coloring agents, one or more flavoring agents, and one or more sweetening agents, such as sucrose or saccharine.

The composition according to the invention can be formulated as oily suspensions by suspending the active ingredient in a vegetable oil, for example, peanut oil, olive oil, sesame seed oil, or coconut oil, or in a mineral oil such as liquid paraffin. The oily suspensions may contain a thickening agent, for example, beeswax, solid paraffin, or cetyl alcohol. Sweetening agents such as those shown above and flavoring agents may be added to provide an agreeable oral composition. These compositions can be preserved by adding an antioxidant such as ascorbic acid.

The composition according to the invention can be formulated in dispersible powder and granule form suitable for the composition of an aqueous suspension by adding water. The active ingredient in such powders and granules is provided in a mixture with a dispersing or wetting agent, suspension agent, and one or more preservatives. The dispersing or wetting agents and suspension agents are exemplified by those already mentioned above. Additional excipients, for example, sweetening agents, flavoring agents, and coloring agents, may also be present.

The composition according to the invention can be in the form of oil in water emulsions. The oil phase can be a vegetable oil, for example, olive oil or peanut oil, or a mineral oil, for example, a liquid paraffin, or a mixture thereof. Suitable emulsifying agents can be natural gums, for example, acacia gum or tragacanth gum, natural phosphatides, for example, soy lecithin, and esters or partial esters derived from fatty acids and hexitol anhydrides, for example, sorbitan monooleate, and products of condensation of partial esters with ethylene oxide, for example, polyoxyethylene sorbitan monooleate. The emulsions may also contain sweetening agents and flavoring agents.

The composition according to the invention can be formulated as syrups and elixirs. The syrups and elixirs can be formulated with sweetening agents, for example, glycerol, propyleneglycol, sorbitol, or sucrose. Such formulations may also contain a demulcent, a preservative, and flavoring agents and coloring agents. Demulcents are protective agents primarily used to alleviate irritation, particularly of mucous membranes or depleted tissues. A number of chemical substances possess demulcent properties. These substances include alginates, mucilages, gums, dextrins, starches, certain sugars, and polymeric polyhydric glycols. Others include acacia gum, agar, benzoin, carbomer, gelatin, glycerin, hydroxyethylcellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose, propyleneglycol, sodium alginate, tragacanth, hydrogels, and the like.

Formulations suitable for oral administration include tablets and coated tablets comprising the compounds of the composition of the invention in a flavored base, such as sucrose, acacia gum, or tragacanth; and pills comprising said compound in an inert base, such as gelatin and glycerin or sucrose and acacia gum.

One skilled in the art will be capable of suitably formulating a liquid formulation, as well as its solid homologues, of the composition of the invention, which contains a suitable amount of each of the compounds of the composition of the invention, depending on the additive or support selected.

As it is used in the present invention, the expression “kit of parts” refers to a product comprising different ingredients, components, or compounds, wherein said ingredients, components, or compounds are physically separated, preferably by separately packaging each ingredient, component, or compound in the kit such that it allows being transported and stored. As will be understood, in the “kit of parts” according to the present invention, the individual active ingredients, components, or compounds represent therapeutic agents and, provided that the use of those compounds, whether simultaneously, separately, or sequentially, produces the new and unexpected therapeutic effect together, as described in the present document, which is not achieved by the compounds independently with respect to one another. Indeed, as demonstrated by the results subsequently, the claimed combination of active ingredients did not represent a mere aggregation of known agents, but rather a new combination with the valuable, surprising property that the combined effect is far more important than the simple sum of the effects that are observed when those active ingredients are used separately. The kit of parts typically comprises its components in suitable containers. The materials suitable for packaging the components of the kit include glass, plastic (polyethylene, polypropylene, polycarbonate, and the like), bottles, vials, paper, pouches, and the like. For example, each container may be in the form of vials, bottles, squeeze bottles, jars, sealed sleeves, sachets or pouches, tubes or blisters or any other suitable form provided that the container is configured to prevent the premature mixing of the components. Each of the different components may be provided separately, or some of the different components may be provided together (i.e., in the same container). Furthermore, the kits of the invention may contain instructions for the simultaneous, sequential, or separate use of the different components that are in the kit. Said instructions may be in the form of printed material or in the form of an electronic medium capable of storing instructions such that they can be read by a subject, such as electronic storage media (magnetic discs, tapes, and the like), optical media (CD-ROM, DVD), and the like. The medium may furthermore or alternatively contain Internet addresses providing said instructions.

It will be understood that the compositions or kits of parts of the invention may comprise, essentially consist, or consist of the aforementioned ingredients, compounds or components.

In this specification the term “comprising” or “comprises” is used to indicate that the composition being described must contain the ingredient(s) listed, but it may optionally contain additional ingredients. The term “essentially consisting of’ or “essentially consists of’ is used to indicate that the composition being described must contain the ingredient(s) listed, and it may also contain a small amount (for example, up to 5 percent by weight, or up to 1 percent by weight, or 0.1 percent by weight) of other ingredients provided that no additional ingredient affects the essential properties of the extract or composition. The term “consisting of’ or “consists of’ is used to indicate that the composition being described must contain only the ingredient(s) listed.

In a particular embodiment, the composition or kit of parts according to the fourth aspect of the invention comprises additional ingredients in addition to hydroxytyrosol and the at least one polyphenol other than hydroxytyrosol. In a particular embodiment, it 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 ingredients in addition to hydroxytyrosol and the at least one polyphenol other than hydroxytyrosol. In another particular embodiment, the hydroxytyrosol and the at least one polyphenol other than hydroxytyrosol comprise 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 ingredients forming the kit.

The term “polyphenol”, as well as the expression “polyphenol that is not hydroxytyrosol wherein the at least one polyphenol other than hydroxytyrosol and/or does not occur naturally in the tree O. europaea” have been defined in the first, second, and third aspects of the invention. Said definitions, as well as the particular embodiments for describing said term and said expression apply to this aspect of the invention.

In a particular embodiment, the hydroxytyrosol is part of a plant extract.

Therefore, in a particular embodiment, and as one skilled in the art will understand, the compound or ingredient “hydroxytyrosol” of the composition and of the kit of parts of the fourth aspect of the invention consists of a plant extract comprising hydroxytyrosol.

The expressions “plant extract”, as well as “is part of a plant extract” have been defined in the first, second, and third aspects of the invention. Therefore, as one skilled in the art will understand, in a particular embodiment, the hydroxytyrosol is supplied in the composition of the fourth aspect of the invention as a plant extract.

In a particular embodiment, the plant extract comprising the hydroxytyrosol of this aspect of the invention is like the extract comprising hydroxytyrosol described in any of the embodiments and definitions of the first, second, and third aspects of the invention.

Therefore, in a particular embodiment, the plant extract comprising hydroxytyrosol is obtained from any plant product of the tree O. europaea, wherein the plant product is any of those indicated in the definition of “plant product” indicated in the first, second, or third aspect of the invention.

In a preferred embodiment of the fourth aspect of the invention, the plant extract of which the hydroxytyrosol is part is an extract of the leaf or of the fruit, preferably the leaf, of the tree O. europaea.

In a particular embodiment, said extract is any extract mentioned in the first, second, and third aspects of the invention, of one or several plant products, preferably of the leaf or of the fruit, more preferably of the leaf, of one or several trees of O. europaea.

In a particular embodiment, the hydroxytyrosol content in the extract comprising hydroxytyrosol of this aspect is any of those indicated in the first, second, and third aspects of the invention for the extract comprising the hydroxytyrosol of said aspects. In a preferred embodiment of the fourth aspect of the invention, the plant extract comprising hydroxytyrosol contains 10-90%, preferably 10% (w/w) of hydroxytyrosol.

In another particular embodiment, the at least one polyphenol that is not hydroxytyrosol of the fourth aspect of the invention is the at least one polyphenol that is not hydroxytyrosol defined and described in the first, second, and third aspects of the invention. Therefore, the polyphenol that is not hydroxytyrosol of the fourth aspect of the invention is any of the polyphenols indicated in the first, second, and third aspects of the invention and defined as belonging to the group of polyphenols that are not hydroxytyrosol.

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.

In another particular embodiment, the at least one polyphenol other than hydroxytyrosol of the group of flavonoids is selected from the group consisting of procyanidins, propelargonidins, prodelphinidins, catechin, epicatechin, quercetin, kaempferol, isorhamnetin, naringin, naringenin, genistein, and rutin.

In a preferred embodiment, the procyanidins are type B procyanidins, preferably selected from the group consisting of type B1 procyanidin, type B2 procyanidin, type B3 procyanidin, type B4 procyanidin, type B5 procyanidin, type B6 procyanidin and type B7 procyanidin. In another particular embodiment, it is a type Cl procyanidin. In a particular embodiment, it is a type A procyanidin, preferably selected from the group consisting of procyanidin Al, procyanidin A2.

In another particular embodiment, the propelargonidins are type A, more preferably type B propelargonidins.

In a particular embodiment, the prodelphinidins are type A.

In another particular embodiment, the at least one polyphenol other than hydroxytyrosol of the group of lignans is selected from the group consisting of secoisolariciresinol diglucoside, secoisolariciresinol, matairesinoside, lariciresinol, pinoresinol, and sesamin In another particular embodiment, the at least one polyphenol other than hydroxytyrosol of the group of stilbenes is selected from the group consisting of resveratrol, piceatannol, pinosylvin, pterostilbene, and rhapontienin, preferably resveratrol.

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.

In another particular embodiment, and as one skilled in the art will understand, the ingredient or compound that is “polyphenol other than hydroxytyrosol” of the composition and kit of parts of the fourth aspect of the invention consists of a plant extract.

In another particular embodiment, the plant extract of which the at least one polyphenol other than hydroxytyrosol is part is not an extract of a plant product of the tree O. europaea , preferably it is not an extract of the leaf or of the fruit of the tree O. europaea.

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

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

In a particular embodiment, the at least one polyphenol other than hydroxytyrosol comprised in each extract comprising a polyphenol other than hydroxytyrosol of this aspect of the invention, is that indicated in the first, second, or third aspect of the invention for said extract.

In a preferred embodiment of the fourth aspect of the invention: - the at least one polyphenol other than hydroxytyrosol which is part of the extract of the skin of the fruit of Prunus dulcis is a flavonoid, preferably selected from the group consisting of procyanidins, propelargonidins, prodelphinidins, catechin, epicatechin, quercetin, kaempferol, isorhamnetin, rutin, and genistein.

- the at least one polyphenol other than hydroxytyrosol which is part of the extract of the fruit of Citrus paradisi or of the extract of the seed of the fruit of Citrus paradisi 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 seed 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 which is part of the extract of the root of Polygonum cuspidatum is a stilbene, preferably selected from the group consisting of resveratrol, piceatannol, pinosylvin, pterostilbene, and rhapontienin, more preferably is resveratrol.

In a particular embodiment, the content and type of flavonoids contained in any extract mentioned in the present invention of one or several plant products, preferably the extract of the skin of the fruit, of one or several trees Prunus dulcis, is that indicated in the first, second, and third aspects of the invention for said extract.

In another particular embodiment, the content and type of flavonoids contained in any extract mentioned in the present invention of one or several plant products, preferably of the fruit or of the seed of the fruit, of one or several trees Citrus paradisi , is that indicated in the first, second, and third aspects of the invention for said extract.

In another particular embodiment, the content and type of lignans contained in any extract mentioned in the present invention of one or several plant products, preferably the extract of the seed of the fruit, of one or several plants of Linum usitatissimum, is that indicated in the first, second, and third aspects of the invention for said extract.

In another particular embodiment, the content and type of stilbenes contained in any extract mentioned in the present invention, of one or several plant products, preferably of the root, of Polygonum cuspidatum, is that indicated in the first, second, and third aspects of the invention for said extract.

Therefore, in a preferred embodiment, the extract of the skin of the fruit of Prunus dulcis contains between 5-60% (w/w), preferably 30% (w/w) of flavonoids, wherein the flavonoids are preferably selected from the group consisting of procyanidins, propelargonidins, prodelphinidins, catechin, epicatechin, quercetin, kaempferol, isorhamnetin, rutin, genistein, and combinations thereof. the extract of the fruit of Citrus paradisi or the extract of the seed of the fruit of Citrus paradisi contains between 10-80% w/w, preferably 45% (w/w) of flavonoids, wherein the flavonoids are preferably selected from the group consisting of naringin, naringenin, quercetin, catechin, rutin, genistein, and combinations thereof. the extract of the seed of the fruit of Linum usitatissimum containing between 5-50% (w/w), preferably 20% (w/w) of lignans, wherein the lignans are preferably selected from the group consisting of secoisolariciresinol diglucoside, secoisolariciresinol, matairesinoside, lariciresinol, pinoresinol, and sesamin, and combinations thereof. the extract of the root of Polygonum cuspidatum contains between 50-100% (w/w), preferably 98% (w/w) of stilbenoids, wherein the stilbenoids are preferably selected from the group consisting of resveratrol, piceatannol, pinosylvin, pterostilbene, rhapontienin, and combinations thereof.

In a particular embodiment, in the composition or kit of parts of the fourth aspect of the invention, the polyphenol other than hydroxytyrosol is at least at 1.25, 1.5, 1.75, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3, 3.1, 3.2, 3.4, 3.5, 3.75, 4, 4.25, 4.5, 4.75, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 10, 11, 12, 51, 18, 20, 21, 22, 23, 24, 24.2, 24.3, 24.4, 24.5, 25, 26, 27, 28, 29, 30, 32, 35, 40, 54, 50, 55, 60, 65, 70, 80, 85, 90, 95, 100, preferably 2.7 times in excess by weight, more preferably 24.3 times in excess by weight, even more preferably at least 28 times in excess by weight with respect to the amount by weight of hydroxytyrosol comprised in the composition or kit of parts.

In another particular embodiment of the fourth aspect of the invention, the w/w ratio of hydroxytyrosokpolyphenol other than hydroxytyrosol is between 1:0.5 and 1:100, 1:1 and 1:50, 1:2 and 1:40, 1:2 and 1:30, 1:2 and 1:28, 1:2 and 1:25, 1:2 and 1:24.5, 1:2 and 1:24.3, 1:2 and 1:24, 1:2 and 1:20, 1:2 and 1: 17, 1:2 and 1:15, 1:2 and 1:10, 1:2 and 1:7, 1:2 and 1:5, 1:2 and 1: 4, 1:2 and 1 :3, preferably between 1:1 and 1:30 in the composition or kit of parts. In another particular embodiment, the w/w ratio of hydroxytyrosokpolyphenol other than hydroxytyrosol 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 in the composition or kit of parts.

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

In a particular embodiment, in the composition or kit of parts of the fourth aspect of the invention, the w/w ratio plant extract comprising hydroxytyrosol: plant extract comprising a polyphenol other than hydroxytyrosol is between 1:0.5 and 1:100, 1:1: and 1:75, 1:2 and 1:50, 1:3 and 1:40, 1:4 and 1:30, 1:5 and 1:25, 1:7 and 1:20, 1:10 and 1: 15, preferably between 1:5 and 1:25, In another particular embodiment, the w/w ratio plant extract comprising hydroxytyrosol: plant extract comprising a polyphenol other than hydroxytyrosol is of 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 of at least 1:10.

In another particular embodiment, the hydroxytyrosol content in the composition of the fourth aspect of the invention is of at least 0.02-20% (w/w), at least 0.025-15% (w/w), at least 0.5- 10% (w/w), at least 0.5-7.5% (w/w), at least 0.6-5% (w/w), at least 0.7-2.5% (w/w), at least 0.8-2% (w/w), at least 0.9-1.5% (w/w), more preferably at least 0.05-10% (w/w), with respect to the total of the composition. In another particular embodiment, the polyphenol composition of almond skin in the composition of the fourth aspect of the invention is of at least 5-100% (w/w), at least 7-75% (w/w), at least 10-50% (w/w), at least 15-40% (w/w), at least 20-30% (w/w), preferably at least 10-50% (w/w) with respect to the total of the composition. In another particular embodiment, the polyphenol composition of almond skin in the composition of the fourth aspect of the invention is of 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), 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) with respect to the total of the composition.

In a preferred embodiment, the content of hydroxytyrosol in the composition of the fourth aspect of the invention is of at least 0.05-10% w/w, preferably at least 0.9% w/w with respect to the total of the composition, wherein at least one polyphenol other than hydroxytyrosol is a polyphenol composition of almond skin, wherein the content of the polyphenol composition of almond skin is of about at least 10-50% w/w, preferably at least 26% w/w with respect to the total of the composition.

In a particular embodiment, the definitions and particular embodiments of the first, second, and third aspects of the invention are applied to the fourth aspect of the invention.

3- Method of the invention

In a fifth aspect, the invention relates to a method for obtaining the composition of the fourth aspect of the invention, which method comprises contacting a composition comprising hydroxytyrosol with a composition comprising at least one polyphenol that is not hydroxytyrosol and that does not occur naturally in the tree O. europaea.

The term “composition” has been defined in the fourth aspect of the invention. The terms “hydroxytyrosol”, and “polyphenol that is not hydroxytyrosol” have been defined in the first, second, and third aspects of the invention.

In a particular embodiment, the polyphenol other than hydroxytyrosol is as defined in any definition and particular embodiment of the first, second, third, and fourth aspects of the invention. As it is used in the present invention, the expression “contacting” refers to the mixture or combination of the composition comprising hydroxytyrosol with the composition comprising a polyphenol other than hydroxytyrosol, such that the ingredients of one composition are surrounded by the ingredients of the other composition. A composition in turn comprising the composition comprising hydroxytyrosol and the composition comprising the polyphenol other than hydroxytyrosol is thus obtained. The product obtained by said contacting is the composition of the first, second, third, or fourth aspect of the invention.

Said contacting can be performed in vitro as described below, or after the composition comprising hydroxytyrosol and the composition comprising a polyphenol other than hydroxytyrosol is ingested by a subject, at the same time or separately. In this case, the contacting between both compositions occurs once they are inside the subject’s body.

The mixture or combination of the composition comprising hydroxytyrosol and the composition comprising a polyphenol other than hydroxytyrosol can be performed by placing in one and the same vessel both compositions and by moving the content of the vessel such that the ingredients of one composition are among the ingredients of the other composition with a certain homogeneity. In a particular embodiment, at least one non-toxic pharmaceutically acceptable excipient suitable for manufacturing the composition of the first, second, third, or fourth aspect of the invention is included in the mixture. Said excipients can be any of the inert diluents, excipients suitable for manufacturing aqueous suspensions, vegetable oils, thickening agents, antioxidants, dispersing or wetting agents, suspension agents, emulsifying agents, sweetening agents, or flavoring agents indicated in the fourth aspect, which the composition of said aspects may comprise.

One skilled in the art will be capable of suitably formulating the mixture, which contains a suitable amount of composition comprising hydroxytyrosol and of composition comprising a polyphenol other than hydroxytyrosol, depending on the excipient or additive selected.

In a particular embodiment, the composition comprising hydroxytyrosol is a plant extract. In another particular embodiment, the composition comprising the at least one polyphenol other than hydroxytyrosol is a plant extract. In a preferred embodiment, the composition comprising hydroxytyrosol is a plant extract and/or the composition comprising the at least one polyphenol that is not hydroxytyrosol is a plant extract.

The plant extract comprising hydroxytyrosol is like the plant extract comprising hydroxytyrosol defined in any of the preceding aspects of the invention.

In a preferred embodiment, the plant extract comprising hydroxytyrosol is an extract of the leaf or of the fruit, preferably of the leaf, of the tree O. europaea.

In a particular embodiment, the hydroxytyrosol content in the extract comprising hydroxytyrosol is any of the hydroxytyrosol content in said extract indicated in any of the preceding aspects of the invention.

In another particular embodiment, the plant extract comprising hydroxytyrosol contains 1- 90% w/w, preferably 10% (w/w) of hydroxytyrosol.

In regard to the plant extract comprising the at least one polyphenol other than hydroxytyrosol is like any extract comprising the at least one polyphenol other than hydroxytyrosol defined in any of the preceding aspects of the invention.

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

In a particular embodiment, the content and type of flavonoids contained in any extract mentioned in the present invention of one or several plant products, preferably the extract of the skin of the fruit, of one or several trees Prunus dulcis is that indicated in the first, second, and third aspects of the invention for said extract. In another particular embodiment, the content and type of flavonoids contained in any extract mentioned in the present invention of one or several plant products, preferably of the fruit or of the seed of the fruit, of one or several trees Citrus paradisi_c is that indicated in the first, second, and third aspects of the invention for said extract.

In particular embodiments: the extract of the skin of the fruit of Prunus dulcis contains between 5-60% w/w, preferably 30% w/w of flavonoids, wherein the flavonoids are preferably selected from the group consisting of procyanidins, propelargonidins, prodelphinidins, catechin, epicatechin, quercetin, kaempferol, isorhamnetin, rutin, genistein, and combinations thereof. the extract of the fruit of Citrus paradisi or the extract of the seed of the fruit of Citrus paradisi contains 10-80% w/w, preferably 45% (w/w) of flavonoids, wherein the flavonoids are preferably selected from the group consisting of naringin, naringenin, quercetin, catechin, rutin, genistein, and combinations thereof. the extract of the seed of the fruit of Linum usitatissimum containing between 5-50% w/w, preferably 20% (w/w) of lignans, wherein the lignans are preferably selected from the group consisting of secoisolariciresinol diglucoside, secoisolariciresinol, matairesinoside, lariciresinol, pinoresinol, and sesamin. the extract of the root of Polygonum cuspidatum contains between 50-100% w/w, preferably 98% (w/w) of stilbenes, wherein the stilbenes are preferably selected from the group consisting of resveratrol, piceatannol, pinosylvin, pterostilbene, rhapontienin, and combinations thereof, preferably is resveratrol, more preferably resveratrol in combination with at least one of the stilbenes indicated previously.

In a particular embodiment, the definitions and particular embodiments of the first, second, third, and fourth aspects of the invention are applied to the fifth aspect of the invention.

4. Pharmaceutical composition of the invention

In a sixth aspect, the invention relates to a pharmaceutical composition comprising the composition of the fourth aspect of the invention, or a composition obtained by the method of the fifth aspect of the invention and a pharmaceutically acceptable excipient.

As used herein, the expression “pharmaceutical composition” refers to any physiologically tolerable composition that typically does not produce an allergic reaction or a similar unfavorable reaction such as gastric disorders, dizziness, or the like, when administered to a human or animal.

Said composition comprises at least one pharmaceutically active ingredient and one or more pharmaceutically acceptable supports. The terms “pharmaceutically acceptable support”, “pharmaceutically acceptable excipient”, “pharmaceutically acceptable diluent”, or “pharmaceutically acceptable vehicle” are used interchangeably herein, and refer to a non toxic filler, diluent, auxiliary or encapsulating material of a solid, semisolid, or liquid formulation of any conventional type. A pharmaceutically acceptable support is essentially non-toxic for receptors at the doses and concentrations used and is compatible with other ingredients of the formulation. Suitable supports include, but are not limited to, water, dextrose, glycerol, saline solution, ethanol, and combinations thereof. The support may contain additional agents such as wettings or emulsifying agents, pH regulating agents, or adjuvants which increase the efficacy of the formulation. The adjuvants could be selected from the group consisting of sterile liquids, such as water and oils, including those from petroleum, animal origin, plant origin, or synthetic origin, such as peanut oil, soybean oil, vaseline oil, sesame seed oil, and the like. Water or aqueous saline solutions and aqueous dextrose and glycerol solutions, particularly for injectable solutions, are preferably used as vehicles. Suitable pharmaceutical vehicles are described in “Remington’s Pharmaceutical Sciences” by E.W. Martin, 21^st Edition, 2005. Except where a conventional support is incompatible with the active ingredient, its use in the therapeutic or pharmaceutical composition is contemplated. Pharmaceutically acceptable excipients also include any of the inert diluents, excipients suitable for manufacturing aqueous suspensions, vegetable oils, thickening agents, antioxidants, dispersing or wetting agents, suspension agents, emulsifying agents, sweetening agents, or flavoring agents indicated in the fourth aspect which the composition of said aspects may comprise.

The pharmaceutical composition comprises a plant extract of any aspect of the invention in a therapeutically effective amount. As it is used herein, the term “effective amount” is synonymous to “therapeutically effective amount”, “effective dose”, or “therapeutically effective dose”, and when used in the present invention it refers to the minimum dose of the plant extract of any aspect of the invention needed to achieve the desired therapeutic effect and includes a sufficient dose to reduce at least one cardiovascular disease symptom, preferably atherosclerosis. The efficacy in treating the diseases or conditions described herein can be determined observing a drop in serum oxLDL levels and oxLDL/LDL ratio. In a particular embodiment, the efficacy in treating the diseases or conditions described herein consists in the description provided in any of the particular embodiments indicated in the first, second, or third aspect of the invention for the expression “increasing the antiatherosclerotic effect of hydroxytyrosol”. In that case, the corresponding reference value can be the indicated in said embodiments or the value of the corresponding parameter in a serum sample from a subject not receiving any type of treatment, or from the same subject to whom the pharmaceutical composition is administered at least 2 weeks before said administration or at least 4 weeks before administration. Therefore, methods for measuring the efficacy in treating the diseases or conditions described herein are also the described in said embodiments of the first, second, or third aspect of the invention.

One skilled in the art can determine a therapeutically effective amount of the inventive compositions when determining the unit dose. As it is used herein, a “unit dose” refers to the amount of inventive composition or kit of parts required to produce a response of the 50 percent of the maximum effect (i.e., ED50). The unit dose can be evaluated by the extrapolation of dosage and response curves derived from system for testing in vitro or in animal models. The amount of compounds in the compositions of the present invention that will be effective in the treatment of a particular disorder or condition will depend on the nature of the disorder or condition and may be determined 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, N.J., 1995; and Drug Facts and Comparisons, Facts and Comparisons, Inc., St. Luis, Mo., 1993). The precise dose to be used in the formulation will also depend on the administration route, and the severity of the disease or disorder, and must be decided according to the medical professional’s assessment and the circumstances of each patient. Several administration patters will be apparent to those skilled in the art.

Furthermore, where the repeated administration of the compounds of any of the preceding aspects of the invention (i.e. hydroxytyrosol, plant extract comprising hydroxytyrosol, polyphenol other than hydroxytyrosol and/or extract comprising a polyphenol other than hydroxytyrosol) is used, an effective amount of said compounds will further depend on factors including, without limitation, the administration frequency, the half-life of the compound, or any combination thereof.

The dosage ranges for the administration of the compositions of the present invention are large enough to produce the desired therapeutic effect. Preferably, the compositions according to the present invention are administered once or twice a day on a regular basis. A typical dose administered to a human is between about 20 mg and about 5 g of the composition, preferably between 200 mg and 1 g of the composition.

The compositions provided herein can be administered to a subject by a number of suitable methods known in the art. Examples of suitable methods include: (1) intramuscular, intradermal, intraepidermal, or subcutaneous administration, (2) oral administration, and (3) topical application (such as ocular, intranasal, and intravaginal application). However, in a preferred embodiment, the compositions are formulated for oral administration.

In some embodiments, the preferred administration route of the compositions provided herein is oral. In those cases, the composition for oral use has the format, form, or formulation, described for the composition of the invention in the fourth aspect of the invention. In a particular embodiment, the definitions and particular embodiments of the first, second, third, fourth, and fifth aspects of the invention are applied to the sixth aspect of the invention.

5. Food and nutritional supplements

As it is used in the present invention, the term “food supplement” or “nutritional supplement” refers to concentrated sources of nutrients or other substances obtained from edible products, the purpose of which fin is to supplement the normal diet. The food supplement or nutritional supplement according to the present invention includes compositions of functional foods, i.e., food, beverage, pet chow or feed, or a food supplement, beverage, pet chow or feed, nutritional supplements, fragrances or flavoring agents, enological or cosmetic formulations. In general, the terms “food supplement” or “nutritional supplement” may also mean:

(i) a product intended to supplement the diet which has or contains one or more of the following nutritional ingredients: [A] a vitamin, [B] a mineral, [C] a herb or other botanical element, [D] an amino acid, [E] a nutritional substance for use by man to supplement the diet by increasing the total nutritional intake; or (F) a concentrate, metabolite, constituent, extract, or combination of any ingredient described in clause (A), (B), (C), (D), or (E); or

(ii) a product which (A) is intended for being ingested; (B) is not represented for use as a conventional food or as a unique product of a meal or the diet; and (C) is labeled as a nutritional supplement.

The “food supplement” or “nutritional supplement” according to the present invention is usually administered orally and is provided together with the diet of a subject. It may be in very different forms, including tablets, capsules, liquid suspensions, dry powder, wet composition, a dry tube feeding or wet tube feeding. It can be provided as a nutritional formulation, for example, medical food, for example, in the form of tube feeding, or in oral nutritional form as a complete meal, as part of a meal, as a food additive, as a powder for dissolution, for example, health drinks, as a solution, as a prepared beverage, including juices, shakes, yogurt drink, smoothie, or soy beverage, in a bar, or dispersed in foods of any type, such as baked goods, cereal bars, dairy bars, fast foods, soups, breakfast cereals, muesli, candies, cookies, cakes.

As it is used in the present invention, the term “nutritionally acceptable excipient” or “nutritionally acceptable support” refers to a support, as it has been defined above, that is edible and relates to preparing solutions to be administered orally. The typical nutritionally acceptable supports, diluents, and excipients will be known to one skilled in the art. Non limiting examples of such supports are provided in US patents 6,258,846, 6,576,666, and 7,112,609.

The amount of the composition present in nutraceutical compositions, nutritional or food products for humans or animals (such as compositions of functional foods, i.e., food, beverage, pet chow or feed, or a food supplement, beverage, pet chow or feed), nutritional supplements, fragrances, or flavoring agents, drugs (pharmaceutical compositions or formulations), veterinary compositions, enological or cosmetic formulations, will vary depending on the application. Typically, the amount of the composition present in the food supplement or nutritional supplement will be from about 0.001 to about 50 percent by weight of the nutraceutical compositions, nutritional or food product, nutritional supplement, fragrance or flavoring agent, enological formulation, such as from about 0.01 to about 10 percent, or from about 0.1 to 1 percent.

In a particular embodiment, the definitions and particular embodiments of the first, second, third, fourth, fifth, and sixth aspects of the invention are applied to the seventh aspect of the invention.

6. Medical uses of the invention

In an eighth aspect, the invention relates to the composition or kit of parts of the fourth aspect of the invention, the composition obtained by the method of the fifth aspect of the invention, the pharmaceutical composition of the sixth aspect of the invention, or the food or nutritional supplement of the seventh aspect of the invention, for use in medicine. In a ninth aspect, the invention relates to the composition or kit of parts of the fourth aspect of the invention, the composition obtained by the method of the fifth aspect of the invention, the pharmaceutical composition of the sixth aspect of the invention, or the food or nutritional supplement of the seventh aspect of the invention, for use in the treatment of a cardiovascular disease.

As it is used in the present invention, the expression “cardiovascular disease” or “CVD” refers to a disease affecting the heart and/or blood vessels. CVD includes coronary artery diseases (CAD) such as angina and myocardial infarction. Other CVDs include atherosclerosis, apoplexy, heart failure, hypertensive heart disease, rheumatic heart disease, cardiomyopathy, abnormal heart rhythms, congenital heart disease, valvular heart disease, carditis, aortic aneurysms, peripheral artery disease, thromboembolic disease, and venous thrombosis. Peripheral vascular disease, cerebrovascular accident, or coronary artery disease are related to atherosclerosis and commonly entail the existence of atherosclerosis.

In a particular embodiment, the cardiovascular disease of the ninth aspect of the invention is selected the group consisting of atherosclerosis, cerebrovascular accident, peripheral vascular disease, and coronary disease.

The term “atherosclerosis” has been defined in the first, second, or third aspect of the invention.

As it is used in the present invention, the expression “peripheral vascular disease” or “PVD” refers to the obstruction of large arteries that are not part of the coronary, aortic arch, or cerebral vasculature. It causes an acute or chronic ischemia (lack of blood supply). EVP can be a consequence of atherosclerosis, inflammatory processes which result in stenosis, an embolism, or the formation of thrombi. Generally, the term EVP is used to refer to atherosclerotic blockages in a lower limb.

In a particular embodiment of the fourth aspect of the invention, the peripheral vascular disease is due to the existence of atherosclerosis.

As it is used in the present invention, the expression “cerebrovascular accident”, “cerebral attack”, “stroke”, or “cerebral infarction” refers to the situation in which the blood flow to a part of the brain is stopped, causing cell death. The “ischemic cerebrovascular accident” or “ischemic stroke”, due to the absence of blood supply, differs from “hemorrhagic cerebrovascular accident” or “ischemic stroke”, due to a hemorrhage. As it is used in the present invention, the expression “ischemic stroke” or “ischemic cardiovascular accident” refers to the situation in which the cerebral structure loses its blood supply due to the sudden and immediate interruption of blood flow due to the occlusion of any of the arteries which supply the encephalic mass. Said occlusion may be due to a buildup of fibrin or calcium or to an erythrocyte abnormality, but it is generally due to atherosclerosis, or due to an embolus (cerebral embolism) that comes from another location, fundamentally the heart or other arteries (such as the carotid or aortic arch bifurcation). As it is used in the present invention, the expression “hemorrhagic stroke” or “hemorrhagic cerebral accident” refers to the situation in which a brain blood vessel ruptures, thereby depriving the area of the brain dependent of this blood vessel, generally an artery, of blood supply. The extravasated blood applies compression on cerebral structures, including other blood vessels, thereby amplifying the area of the brain affected.

In a particular embodiment of the ninth aspect of the invention, the cerebrovascular accident is ischemic, preferably due to the existence of atherosclerosis.

As it is used in the present invention, the expression “coronary disease” or “coronary artery disease” refers to the condition in which there is an imbalance between the blood flow of the coronary arteries or coronary flow and the oxygen requirement of the heart muscle or myocardium. This imbalance causes ischemia the effects of which are metabolic (increase in lactic acid, acidosis, decrease in ATP, decrease in phosphocreatines), mechanical (decrease in cardiac contractility, decrease in distensibility of the ischemic area, and others) and electrical (modification of rest and action potentials, electric instability and the subsequent rhythm disorders). The main cause of coronary disease is the narrowing of the coronary arteries supplying blood to the heart due to atherosclerosis.

In a particular embodiment of the fourth aspect of the invention, the coronary disease is due to the existence of atherosclerosis.

Depending on the disorder and on the subject to be treated, as well as the administration route, the compositions, components of the kit of parts, pharmaceutical compositions, food or nutritional supplements of the invention can be administered at variable doses (i.e., therapeutically effective doses, administered to a patient in need of same). In this regard, one skilled in the art will appreciate that the dose administered to a mammal, particularly a human, in the context of the present invention must be sufficient so as to affect a therapeutic response in the mammal during a reasonable space of time. In a particular embodiment, said therapeutic effect corresponds with the efficacy in treating the diseases or conditions described herein specified in the sixth aspect of the present invention. One skilled in the art will recognize that the selection of the dose and exact composition and the most suitable dosage regimen will also be influenced by, among others, the pharmacological properties of the formulation, the nature and severity of the condition being treated, and the physical condition of the receptor, as well as the age, condition, body weight, sex, and response of the patient to be treated, and the stage/severity of the disease.

As it is used in the present invention, the term “subject” refers to a mammal, preferably a human.

The subject to be treated is a mammal, preferably a human. The subject to be treated according to the invention can be selected based on several criteria associated with cardiovascular diseases, such as serum oxLDL and/or LDL levels, or the corresponding oxLDL/LDL- cholesterol ratio.

In a particular embodiment, the subject presents hypercholesterolemia, preferably moderate hypercholesterolemia. In another particular embodiment, the subject presents acute hypercholesterolemia.

Therefore, in a preferred embodiment, the composition, the kit of parts, the pharmaceutical composition, or the food or nutritional supplement for use of the eighth or ninth aspect of the invention is administered to a patient with hypercholesterolemia, preferably moderate hypercholesterolemia.

As it is used in the present invention, the term “hypercholesterolemia” refers to the condition characterized by high blood cholesterol levels. Given that cholesterol is insoluble in water, it is transported in blood plasma together with proteins, forming lipoproteins. Lipoproteins are classified by their density: very low-density lipoproteins (VLDL), intermediate-density lipoproteins (IDL); low-density lipoproteins (LDL), and high-density lipoproteins (HDL). All lipoproteins transport cholesterol, however high levels of lipoproteins other than HDL, particularly high levels of LDL transporting cholesterol (LDL-cholesterol), tend to be deposited on arterial walls, forming atheroma plaques and favoring the development of atherosclerosis and coronary disease, stroke, and peripheral arterial disease. Generally, cholesterol levels are considered to be high when the concentration of total blood cholesterol is greater than 200 mg/dl, and/or the concentration of LDL-cholesterol is greater than 100 mg/dl.

As it is used in the present invention, the expression “moderate hypercholesterolemia” refers to hypercholesterolemia characterized by a concentration of total blood cholesterol greater than 200 mg/dl and less than 250 mg/dl, and a concentration of LDL-cholesterol greater than 100 mg/dl and less than 175 mg/dl. As it is used in the present invention, the expression “acute hypercholesterolemia” refers to hypercholesterolemia characterized by a concentration of total blood cholesterol greater than 250 mg/dl, and a concentration of LDL-cholesterol greater than 175 mg/dl.

While the compositions, components of the kit of parts, the pharmaceutical composition, or the food or nutritional supplement of the invention can be administered as such, the invention also contemplates the possibility of at least one of the components of the composition, kit of parts, pharmaceutical composition, or food or nutritional supplement to be administered separately from the rest of the components of the composition, of the kit of parts, of the pharmaceutical product, or of the food or nutritional supplement.

Therefore, in a particular embodiment of the eighth and ninth aspects of the invention, the hydroxytyrosol and the polyphenols other than hydroxytyrosol are administered separately.

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

In another particular embodiment, the at least one polyphenol other than hydroxytyrosol is administered separately from the hydroxytyrosol or from the extract comprising the hydroxytyrosol. In another particular embodiment, the at least one polyphenol other than hydroxytyrosol is administered separately from the hydroxytyrosol or from the extract comprising the hydroxytyrosol and from the rest of the components of the composition, kit of parts, pharmaceutical composition, or food or nutritional supplement of the invention.

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

In a particular embodiment, the different parts of the kit for use of the eighth and ninth aspects of the invention can be administered separately or can alternatively be combined before the administration. In a preferred embodiment of the eighth and ninth aspects of the invention, the components of the kit of parts are combined before the administration.

In an embodiment of the eighth and ninth aspects of the invention, the administration of the composition, the components of the kit of parts, the pharmaceutical product, or the food or nutritional supplement comprises the administration of multiple doses of the composition, of the components of the kit of parts, of the pharmaceutical product, or of the food or nutritional supplement of the fourth aspect of the invention, 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 for at least 1 week, more preferably at least 2 weeks, even more preferably for at least 4 weeks or for at least 8 weeks.

In another embodiment, the administration of the composition, the components of the kit of parts, the pharmaceutical product, or the food or nutritional supplement comprises the administration of one, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 15, 20, preferably 2 daily doses of the composition, of the components of the kit of parts, of the pharmaceutical product, or of the food or nutritional supplement of the invention.

Those skilled in the art will recognize that the initial indications of the suitable therapeutic dose of the compositions of the invention can be determined in vitro and in animal model systems in vivo , and in clinical trials in humans. One skilled in the art would know how to use animal studies and human experience to identify a dose that can be administered safely without generating toxicity or other side effects. For acute treatment where it is desirable to substantially restore serum oxLDL levels, the serum oxLDL/LDL ratio, the therapeutic dose is preferably close to the maximum tolerated dose. For chronic preventive use, lower doses may be desirable due to concerns for long-term effects.

In another embodiment of the eighth and ninth aspects of the invention, the administration of the composition, of the components comprised in the kit of parts, of the pharmaceutical composition, or of the food or nutritional supplement comprises administering 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, preferably 7.5 mg/day of hydroxytyrosol.

In another particular embodiment of the eighth and ninth aspects of the invention, the administration of the composition, of the components comprised in the kit of parts, of the pharmaceutical composition, or of the food or nutritional supplement comprises administering 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 of polyphenols other than hydroxytyrosol.

In another particular embodiment of the eighth and ninth aspects of the invention, the administration of the composition, of the components comprised in the kit of parts, of the pharmaceutical composition, or of the food or nutritional supplement comprises administering 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, preferably 7.5 mg/day of hydroxytyrosol and any of the doses in mg/day of polyphenols other than hydroxytyrosol indicated above, preferably 210 mg/day of polyphenols other than hydroxytyrosol.

Therefore, in a particular embodiment of the eighth and ninth aspects of the invention, the administration of the composition, of the components comprised in the kit of parts, of the pharmaceutical product, or of the food or nutritional supplement of the invention comprises administering 7.5 mg/day of hydroxytyrosol and 210 mg/day of polyphenols other than hydroxytyrosol.

In another particular embodiment of the eighth and ninth aspects of the invention, the administration of the composition, of the components comprised in the kit of parts, of the pharmaceutical composition, or of the food or nutritional supplement comprises administering 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, 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 of plant extract providing hydroxytyrosol as defined in the first to fourth aspects of the invention. In another particular embodiment of the eighth and ninth aspects of the invention, the administration of the composition, of the components comprised in the kit of parts, of the pharmaceutical composition, or of the food or nutritional supplement comprises administering 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, 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 of plant extract providing polyphenols as defined in the first to fourth aspects of the invention.

In another particular embodiment of the eighth and ninth aspects of the invention, the administration of the composition, of the components comprised in the kit of parts, of the pharmaceutical composition, or of the food or nutritional supplement, comprises administering 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, 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 of plant extract providing hydroxytyrosol as defined in the first to fourth aspects of the invention and any of the doses indicated above in mg/day of plant extract providing polyphenols as defined in the first to fourth aspects of the invention, preferably 700 mg/day.

In a preferred embodiment of the eighth and ninth aspects of the invention, the administration of the composition, of the components comprised in the kit of parts, of the pharmaceutical composition, or of the food or nutritional supplement, comprises administering 67 mg/day of plant extract providing hydroxytyrosol as defined in the first to fourth aspects of the invention and 700 mg/day of plant extract providing polyphenols in the first to fourth aspects of the invention. In a particular embodiment, the definitions and particular embodiments of the first, second, third, fourth, fifth, sixth, and seventh aspect of the invention are applied to the eighth and ninth aspects of the invention.

In a particular embodiment, the term consists of, essentially consists of, and comprises are interchangeable in any embodiment of any aspect of the present invention.

The invention is described below by means of the following merely illustrative and non limiting examples of the scope of the invention.

EXAMPLES Materials and methods

A- Materials and methods corresponding to Examples 1 and 2

Study subjects

Volunteers were recruited at the Endocrinology Department of Hospital San Cecilio of Granada (Spain) and from the database of the clinical studies unit of Biosearch, S.A. The participants in the study met the following inclusion criteria: men and women 18 to 65 years of age, with moderate hypercholesterolemia (LDL-col >100 mg/dL and/or total cholesterol >200 mg/dL) and who are not receiving medical treatment for this condition. The exclusion criteria included pregnancy, having an important medical issue, diabetes, and/or neurovascular disease, and presenting an allergy to the antibiotics or to any of the ingredients of the study. The subjects were also excluded if they were on a diet or taking food supplements during the study. The study was carried out according to the Declaration of Helsinki, and the protocol was approved by the Regional Ethics Committee (Granada, Spain). The informed consent was obtained from all the subjects. The trial was recorded in the US Library of Medicine (http://www.clinicaltrial.gov) as NCT04029727.

About 300 subjects were contacted and invited to participate in the study, and the suitability of 63 candidates was evaluated. Out of those 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 intervention and were randomly assigned to the study groups (Figure 1).

Design of the study It was a randomized, parallel, double-blind, placebo-controlled pilot study with an 8-week duration. The study was performed between May and July of 2019. 32 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 (7.5 mg HT + 210 mg ASPs + 33 mg maltodextrin per day), and the other group (control group) received a placebo (800 mg maltodextrin per day). The treatments were given to the subjects for 8 weeks and they took 2 capsules per day (taking the supplement with extracts or the placebo) at lunch. The participating subjects were instructed not to deviate from their regular habits and to maintain their normal diet and physical activity over the 8 weeks. Neither the researchers nor the subjects knew what treatment sequence the subjects had been assigned to; the researchers did not learn this information until the end of the study.

Products studied

Each capsule with extract contained 3.75 mg of HT in 33.5 mg of an extract of olive tree ( Olea europaea L.), and 105 mg of ASP in 350 mg of an extract of almond skin ( Prunus dulcis (Mill.)) and 16.5 mg of maltodextrin. Each capsule with placebo contained 400 mg of maltodextrin. Both extracts were obtained in the Biosearch, S.A. production plant in Talayuela, Caceres (Spain) and the combination of the extracts was performed at the Biosearch, S.A. R&D facilities in Granada (Spain). The capsules were prepared in the Pharmaceutical Technology Department of the Faculty of Pharmacy of the University of Granada (Spain). The extracts and placebo were supplied in identical gelatin capsules, packaged in identical plastic containers, with a code number that referred to the code of the volunteer according to randomization.

Preparation of the plant extracts

The olive tree extract can be obtained by means of methods based on the extraction of leaves of O. europaea L. with an aqueous solvent, such as water for example. The method includes the following steps:

(i) extracting leaves of O. europaea with an aqueous solvent at a temperature of about 40 a 100°C for 1-10 hours

(ii) treating the extract obtained in step (i) with activated carbon and then removing the activated carbon. The extract of Prunus dulcis Mill can be obtained following the same method as that used for obtaining the extract of O. europaea L., wherein the starting material is the skin of the fruit instead of leaves of O. europaea.

Optionally, step (i) may include several extraction phases for extracting the plant product with the initial solvent or a different solvent, combining the extracts obtained to continue the process. Alternatively, the extract may also be concentrated by means of chromatography using polymeric adsorbent resins. The method can also include a final drying step for drying the extract obtained in the preceding steps of the method.

Parameters of the study and obtaining data

The primary endpoint of the study was the determination of oxLDL levels. The 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.

The subjects went to the Biosearch, S.A. facilities in Granada at the start of the study and at 4 and 8 weeks. Blood samples were collected after an overnight fast that lasted at least 10 hours, before the start of the intervention and in each follow-up visit, using the Vacutainer® SST™ II Advance Tubes (BD,NJ, USA) system containing a thixotropic gel. The serum was obtained after centrifugation of the blood samples at 1000 g for 15 min and stored at -80°C.

Anthropometric measurements were taken in each visit using standard methods. The weight was determined using the Tanita BC-418 body composition analyzer (Tanita, Tokyo, Japan). The height was determined using a height measuring apparatus with a precision of 1 mm (range, 80-200 cm). The body mass index (BMI) was calculated as weight/height squared (kg/m2). All the measurements were performed by qualified personnel.

The food intake of each subject was evaluated at the start and at the end of the study to control possible changes in their habits. The subjects completed a report detailing 72 hours of their dietary intake, specifying the types of foods consumed and the weights of the portions. The daily food, the intake of energy, the intake of nutrients, and the energy provided by the macronutrients were calculated using the Nutriber computer application (Funiber, Barcelona, Spain). Subjects were also asked about the physical activity habits (type of activity and time they practice it per week) at the start and at the end of the study.

Compliance with the protocol in regard to the consumption of the product was verified at the end of the intervention by comparing the number of capsules supplied and the number of capsules returned. Adverse events, defined as any unfavorable and unwanted effect, were recorded in the follow-up visits (at four and eight weeks).

Analysis of biochemical parameters

The levels of the lipid parameters (TC, LDL-c, HDL-c, and TG) were analyzed by an external laboratory (Reference Laboratory S.A, Barcelona, Spain) by means of standard methods. The determination of serum oxLDL concentration was performed by means of ELISA specific for human oxLDL (Elabscience Biotechnology, USA); and the oxLDL/LDL-c ratio was calculated.

Statistical analysis

The normal distribution of all the variables was verified by means of normal probability graphs and the Shapiro-Wilk test. The data was presented as mean (and SD) for continuous variables and as n (%) for categorical variables.

For comparisons between groups at the start of the study (extract vs. control), the continuous variables were analyzed with the Student’s t-test or with the Kruskall- Wallis non-parametric method, as appropriate, while the categorical variables were analyzed with the Chi-squared test.

All the parameters between groups were compared and the variation with respect to the initial intra-group values was also analyzed. To that end, bivariate analysis, as well as a fit test based on mixed regression models will be performed. The comparison between the means of the experimental group and the control group was performed by means of the T-test when normality could be assumed. When normality could not be assumed, the non-parametric U- Mann Whitney test was performed. Furthermore, the mixed linear regression model, which represents repeated measurements throughout the study (intra- subject-random effect), was fitted to the responses in order to verify changes over time and between groups, as well as to find significant factors associated with the responses. A general alpha level of 0.05 was used as the cut-off point for statistical significance. The statistical analysis was carried out using SPSS version 26.0 software for Windows (SPSS, Chicago, IL, USA).

B- Materials and methods corresponding to Examples 3 and 4

Reagents a-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)

Basic medium (“ Medium used to feed the reactor SHI MIC in Molly K. et al. Microbial Ecology in Health and Disease 7:191-200)

Plant extracts

Table 1. Plant extracts used in this study The olive tree and almond extract coincide with those used in section A on materials and methods. Therefore, they may also be obtained using the methods indicated in said section.

The grapefruit, flax, and polygonum extracts can be obtained by applying the same method for obtaining the olive tree extract indicated in section A- on materials and methods, wherein:

- the starting material for the grapefruit extract is the seed of the fruit of the tree Citrus paradisi MacFad

- the starting material for the flax extract is the seed of the fruit of Linum usitatissimum L. - the starting material for the polygonum extract is the root of Polygonum cuspidatum Sieb. etZucc.

Moreover, the solvent used in the methods for obtaining these extracts is a hydroalcohol, wherein the alcohol can be ethanol or methanol, for example.

Chromatographic analysis

Solutions of each of the extracts were resolved by means of chromatography in an Agilent 1200s equipment with a binary pump and DAD SL detector; with stationary phase Phenomenex Luna C18 5 pm 250x4.6 mm at 30 °C and mobile phase: A: 1% formic acid (v/v); B: acetonitrile:methanol 80:20 (flow: 0.8-1 mL/min). 10 pL of sample were injected, and for the run (65 min) absorbance at 330 and 280 nm was measured. The chromatographic conditions, with respect to the composition of the mobile phase and flow, are summarized in the following table. Table 2. Chromatographic conditions

In vitro digestion on a small scale

The simulation of human (adult) digestion on a small scale was carried out at room temperature, in 4 semicontinuously connected compartments, with the characteristics indicated in Table 3.

Table 3. Compartments simulated in the digestion process on a small scale

The steps of the digestion process, as well as the transfers of sample between compartments for treating or simulating a dynamic process, are summarized in Table 4.

Table 4. Digestion process on a small scale ^he volume (mL) contained in each compartment at all times (minutes), as well as the ²volume transferred between them: M-S, mouth to stomach; S-SI, stomach to small intestine; SI-C, small intestine to colon, is indicated. 3 Volume of acid and base added to the stomach and small intestine, respectively, to modify the pH from 4 (initial stomach) to 2 (final stomach) and maintain a value close to 6.5 (small intestine). The time interval during which digestion takes place in each compartment is highlighted with bold and underlining.

Example 1. Effect of the combination of extracts of standard olive tree ( Olea europaea L.) and skin of a standard almond ( Prunus dulcis Mill.) on serum oxLDL levels

A total of 30 volunteers completed the study, since two subjects of the extract group refused to participate before attending the initial visit (Figure 1). It was confirmed that the compliance rate was very high (about 100%). No adverse events resulting from the intake of either of the two types of treatment capsules were reported. Of the 30 volunteers who finished the study, 13 were men (43.43%) and 17 were women (56.7%). The mean age of the subjects was 48.13±12.9 years, and they presented a mean BMI of 25.38±3.33 kg/m2. No significant differences in the baseline characteristics of the volunteers were detected (Table 5).

Table 5. Initial characteristics of the subjects participating in the study

Mean ± standard deviation (SD) for continuous variables and n (%) for categorical variables. P indicates differences between groups.

Table 6 shows the intake of energy and nutrients at the baseline and at week 8. There were no significant differences at the baseline between intervention groups, or between the baseline and the end of the intervention in each (extract or control) group. Nor were there any changes during the study in the degree of physical activity (data not shown) and the subjects in both groups did not significantly change their BMI between the start and week 8 (Table 6).

8 weeks. Means ± SD. P indicates differences between initial values and values after 8 weeks of treatment, in each of the groups. %En: value expressed as % of total energy.

5 Table 7 shows the values of the parameters of lipids and the variables of oxidized lipids evaluated for the study. No significant changes between the baseline and the end of the intervention in serum TC, LDL-c, HDL-c, TG were observed in any group. In regard to the state of oxidation of the lipids, oxLDL levels increased significantly from the baseline in the control group at week 4 (p=0.01) and at week 8 (p=0.03), while they tended to decrease in the 10 extract group, being significantly lower in the extract group than in the control group after 8 weeks of treatment (p<0.001). When the oxLDL/LDL-c ratio is analyzed, a precise estimate of oxidation in vivo of LDL(19), a significant increase in the oxLDL/LDL-c ratio from the baseline in the control group is observed at week 4 (p=0.001) and at week 8 (p=0.002), and a significant decrease of this proportion in the extract group is observed at week 8 (p=0.047), 15 the levels of the oxLDL/LDL-c ratio in the extract group also being significantly lower than in the control group after 8 weeks of treatment (p<0.001) (Table 7 and Figure 2).

Table 7. Lipid and oxidized LDL parameters of the subjects who took the placebo or the extract, at the start, at 4 weeks, and at 8 weeks of treatment. Mean ± SD adjusted for sex, age, obesity/overweight, smoker, and physical activity. P indicates differences between groups at each time. * P <0.05 between start, 4 weeks and 8 weeks

Example 2. Effect of mixtures of olive tree, almond, grapefruit, flax, and polygonum extract on the bioavailability of hydroxytyrosol of the olive tree extract. To verify if the presence of other polyphenols favors the bioavailability of hydroxytyrosol present in the olive tree extract, digestion assays were performed on a small scale on the extracts separately and in different combinations. As a reference, 50 mg/mL olive tree extract and 1:1, 1:4, and 1:9 mixtures with almond skin extract were used. Based on the ratio of polyphenols of both extracts in each of the mixtures, the mixtures with the 3 other assayed extracts were defined (Table 9).

Table 9. Mixtures of extracts, their initial concentration in mouth (mg/mL) and the polyphenol concentration (mg/mL)

First, an assay was carried out with the mixtures of olive tree and almond extract from which the content of hydroxytyrosol of the final small intestine samples (370 min of digestion) was analyzed by means of HPLC-DAD, followed by an interpolation of the data obtained from a hydroxytyrosol calibration line. Table 10 shows the results (ppm) of said analysis.

Table 10. Concentration (ppm) of HT in intestinal and colonic samples in vitro (small scale) and % of the theoretical maximum derived from the extract dose and dilution factor. The results of this assay first of all seem to indicate that the HT present in the olive tree extract is degraded by about 50% after the digestion process in the compartment simulating the small intestine, under the present assay conditions. Moreover, the presence of other polyphenols allows the concentration of HT to increase, and it does so exponentially. Under these experimental conditions, the maximum concentration of almond could allow close to 90% of the HT consumed to be available in the intestinal lumen for absorption.

Given that the in vivo absorption of HT presumably occurs in the small intestine, the presence of this phenolic acid was analyzed only in the intestinal samples (370 min of digestion) obtained from the digestion assays on a small scale performed with the mixtures of olive tree extract and the other extracts (Table 11).

Table 11. Concentration (ppm) of HT in in vitro intestinal samples (small scale) and % of the theoretical maximum derived from the extract dose and dilution factor (340.2 ppm).

The results obtained indicate that the presence of other polyphenols, regardless of their structure or origin, provide a degree of protection to the hydroxytyrosol of the olive tree extract equivalent to that detected in the case of the polyphenols of the almond extract.

Example 4. Analysis of the polyphenols present in each of the plant extracts

In regard to the characterization of the plant extracts, the method for polyphenols analysis by means of high-performance liquid chromatography (HPLC) was developed internally by Biosearch, S.A. for the qualitative analysis of polyphenols of plant extracts and complex mixtures, being able to classify them in some cases by families of polyphenols based on the retention time obtained; or for the quantitative analysis of specific polyphenols, combining the previous information with the response of the corresponding chromatographic standard.

Although there are other methods for the determination of polyphenols by HPLC, the advantage of the method used allows the different families and types of polyphenols to be resolved in the same chromatogram, maximally reducing overlap between signals, which could accordingly hinder their identification. This efficacy in regard to the separation of signals is achieved as a result of the (previously indicated) chromatographic conditions of the method. A 65-minute chromatogram is obtained under these conditions, which allows, as previously indicated, the different signals corresponding to polyphenols of a sample to be resolved without overlap between the different families of polyphenols. A DAD detector is used, setting the detection for signals with the usual wavelengths for the detection of polyphenols: 280 and 330 nm.

The chromatographic assays performed allow demonstrating that the hydroxytyrosol present in the olive tree fruit extract is not present in the other evaluated samples containing other types of polyphenols (peak detected at 280 nm and an approximate retention time (RT) of 10.5 minutes; Figures 3 to 6). Likewise, the group of major compounds detected in the almond extract, both at 280 and at 330 nm, with RTs greater than 35 min, are not present in the olive tree extract or their representation within the components of this extract can be considered residual (Figure 3). Upon joint analysis of the olive tree and grapefruit extract, the results are similar: the main components of each of the extracts (hydroxytyrosol with an RT of 10.5 min in olive tree, and flavonoids with an RT between 35 and 45 min in grapefruit) are not detected in the other extract (Figure 4). Upon comparison of the polyphenol composition of the olive tree and flax extract, it is again verified that compounds such as lignans, with an RT greater than 40 min, are not detected for the most part in the olive tree extract (Figure 5). Finally, the polygonum extract is characterized by a high concentration of the stilbene resveratrol, with an approximate RT of 43 minutes, without the presence of compounds with an RT of 10-11 min (characteristic of hydroxytyrosol) and without this type of compounds being detected significantly in the present chromatograms in the olive tree extract (Figure 6).

Based on these results, it can be affirmed that the hydroxytyrosol present in the olive tree fruit extract is not present in the other samples evaluated containing other types of polyphenols, and likewise the flavonoids, lignans, and stilbenes characterizing the almond, grapefruit, flax, and polygonum extracts are not found significantly in the olive tree extract.

Claims

1. Use of at least one polyphenol that is not hydroxytyrosol and/or that does not occur naturally in the tree Olea europaea (O. europaea) for increasing the bioavailability of hydroxytyrosol after the oral administration of hydroxytyrosol or for increasing the antiatherosclerotic effect of hydroxytyrosol after the oral administration of hydroxytyrosol.

2. A polyphenol that is not hydroxytyrosol and/or that does not occur naturally in the tree Olea europaea (O. europaea) for use in increasing the bioavailability of hydroxytyrosol after the oral administration of hydroxytyrosol or in increasing the antiatherosclerotic activity of hydroxytyrosol after the oral administration of hydroxytyrosol.

3. The hydroxytyrosol for use in the treatment of atherosclerosis, wherein hydroxytyrosol is administered orally in combination with a polyphenol that is not hydroxytyrosol and/or that does not occur naturally in the tree Olea europaea (O. europaea).

4. The use according to claim 1, the polyphenol for use according to claim 2, or the hydroxytyrosol for use according to claim 3, wherein the polyphenol that is not hydroxytyrosol is administered orally.

5. The use according to any of claims 1 or 4, the polyphenol for use according to any of claims 2 or 4, or the hydroxytyrosol for use according to any of claims 3 or 4, wherein the hydroxytyrosol is part of a plant extract.

6. The use, the polyphenol for use, or the hydroxytyrosol for use according to claim 5, wherein the plant extract of which the hydroxytyrosol is part is an extract of the leaf or of the fruit of the tree Olea europaea.

7. The use, the polyphenol for use, or the hydroxytyrosol for use according to any of claims 5 or 6, wherein the plant extract comprising hydroxytyrosol contains 1-90% w/w, preferably 10% (w/w) of this compound.

8. The use according to any of claims 1, 4-7, the polyphenol for use according to any of claims 2, 4-7, or the hydroxytyrosol for use according to any of claims 3-7, wherein the at least one polyphenol is selected from the group of polyphenols consisting of flavonoids, lignans, and stilbenes.

9. The use, the polyphenol for use, or the hydroxytyrosol for use according to claim 8, wherein the at least one polyphenol of the group of flavonoids is selected from the group consisting of procyanidins, propelargonidins, prodelphinidins, catechin, epicatechin, quercetin, kaempferol, isorhamnetin, naringin, naringenin, genistein, and rutin.

10. The use, the polyphenol for use, or the hydroxytyrosol for use according to claim 8, wherein the at least one polyphenol of the group of lignans is selected from the group consisting of secoisolariciresinol diglucoside, secoisolariciresinol, matairesinoside, lariciresinol, pinoresinol, and sesamin.

11. The use, the polyphenol for use, or the hydroxytyrosol for use according to claim 8, wherein the at least one polyphenol of the group of the stilbenes is selected from the group consisting of resveratrol, piceatannol, pinosylvin, pterostilbene, and rhapontienin.

12. The use according to any of claims 1, 4-11, the polyphenol for use according to any of claims 2, 4-11, or the hydroxytyrosol for use according to any of claims 3-11, wherein the at least one polyphenol is part of a plant extract.

13. The use, the polyphenol for use, or the hydroxytyrosol for use according to claim 12, wherein the plant extract of which the polyphenol is part is not an extract of the leaf or of the fruit of the tree Olea europaea.

14. The use, the polyphenol for use, or the hydroxytyrosol for use according to any of claims 12 or 13, wherein the plant extract of which the at least one polyphenol is part is an extract selected from the group consisting of

- an extract of the skin of the fruit of Prunus dulcis - an extract of the fruit of Citrus paradisi or an extract of the seed of the fruit of Citrus paradisi

- an extract is of the seed of the fruit of Linum usitatissimum, and

- an extract of the root of Polygonum cuspidatum.

15. The use, the polyphenol for use, or the hydroxytyrosol for use according to claim 14 wherein:

- the at least one polyphenol other than hydroxytyrosol which is part of the extract of the skin of the fruit of Prunus dulcis is a flavonoid, preferably selected from the group consisting of procyanidins, propelargonidins, prodelphinidins, catechin, epicatechin, quercetin, kaempferol, isorhamnetin, rutin, and geni stein,

- the at least one polyphenol other than hydroxytyrosol which is part of the extract of the fruit of Citrus paradisi or the extract of the seed of the fruit of Citrus paradisi is a flavonoid, preferably selected from the group consisting of naringin, naringenin, quercetin, catechin, rutin, and geni stein,

- the at least one polyphenol other than hydroxytyrosol which is part of the extract of the seed 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 which is part of the extract of the root of Polygonum cuspidatum is a stilbene, preferably selected from the group consisting of resveratrol, piceatannol, pinosylvin, pterostilbene, and rhapontienin.

16. The use according to any of claims 14 or 15, wherein:

- the extract of the skin of the fruit of Prunus dulcis contains between 5-60% (w/w), preferably 30% (w/w) of flavonoids, wherein the flavonoids are preferably selected from the group consisting of procyanidins, propelargonidins, prodelphinidins, catechin, epicatechin, quercetin, kaempferol, isorhamnetin, rutin, genistein, and combinations thereof,

- the extract of the fruit of Citrus paradisi or the extract of the seed of the fruit of Citrus paradisi contains between 10-80% w/w, preferably 45% (w/w) of flavonoids, wherein the flavonoids are preferably selected from the group consisting of naringin, naringenin, quercetin, catechin, rutin, geni stein, and combinations thereof,

- the extract of the seed of the fruit of Linum usitatissimum containing between 5-50% (w/w), preferably 20% (w/w) of lignans, wherein the lignans are preferably selected from the group consisting of secoisolariciresinol diglucoside, secoisolariciresinol, matairesinoside, lariciresinol, pinoresinol, and sesamin, and combinations thereof,

- the extract of the root of Polygonum cuspidatum contains between 50-100% (w/w), preferably a 98% (w/w) of stilbenes, wherein the stilbenes are preferably selected from the group consisting of resveratrol, piceatannol, pinosylvin, pterostilbene, rhapontienin, and combinations thereof.

17. The use according to any of claims 1, 4-16, the polyphenol for use according to any of claims 2, 4-16, or the hydroxytyrosol for use according to any of claims 3-16, wherein the polyphenol is used at least 2.7 times in excess by weight with respect to the amount by weight of hydroxytyrosol orally administered.

18. A composition or kit of parts comprising hydroxytyrosol and at least one polyphenol, wherein the at least one polyphenol is not hydroxytyrosol and/or that does not occur naturally in the tree O. europaea.

19. The composition or kit of parts according to claim 18, wherein the hydroxytyrosol is part of a plant extract.

20. The composition or kit of parts according to claim 19, wherein the plant extract of which the hydroxytyrosol is part is an extract of the leaf or of the fruit of the tree of O. europaea.

21. The composition or kit of parts according to claim 20, wherein the plant extract comprising hydroxytyrosol contains 1-90% w/w, preferably 10% (w/w) of this compound.

22. The composition according to claims 18 to 21, wherein the at least one polyphenol is selected from the group consisting of flavonoids, lignans, and stilbenes.

23. The composition according to claim 22, wherein the at least one polyphenol of the group of flavonoids is selected from the group consisting of procyanidins, propelargonidins, prodelphinidins, catechin, epicatechin, quercetin, kaempferol, isorhamnetin, naringin, naringenin, geni stein and rutin.

24. The composition according to claim 22, wherein the at least one polyphenol of the group of lignans is selected from the group consisting of secoisolariciresinol diglucoside, matairesinoside, lariciresinol, pinoresinol, sesamin, and secoisolariciresinol.

25. The composition according to claim 22, wherein the polyphenol of the group of the stilbenes is selected from the group consisting of resveratrol, piceatannol, pinosylvin, pterostilbene, and rhapontienin.

26. The composition or kit of parts according to any of claims 18-25, wherein the at least one polyphenol is part of a plant extract.

27. The composition or kit of parts according to claim 26, wherein the plant extract of which the polyphenol is part is not an extract of the leaf or of the fruit of the tree of O. europaea.

28. The composition or kit of parts according to any of claims 26 or 27, wherein the plant extract of which the at least one polyphenol is part is an extract selected from the group consisting of:

- an extract of the skin of the fruit of Prunus dulcis.

- an extract of the fruit of Citrus paradisi or an extract of the seed of the fruit of Citrus paradisi

- an extract of the seed of the fruit of Linum usitatissimum

- an extract of the root of Polygonum cuspidatum.

29. The composition or kit of parts according to claim 28, wherein: - the at least one polyphenol which is part of the extract of the skin of the fruit of Primus dulcis is a flavonoid, preferably selected from the group consisting of procyanidins, propelargonidins, prodelphinidins, catechin, epicatechin, quercetin, kaempferol, isorhamnetin, rutin, and genistein,

- the at least one polyphenol which is part of the extract of the fruit of Citrus paradisi or the extract of the seed of the fruit of Citrus paradisi is a flavonoid, preferably selected from the group consisting of naringin, naringenin, quercetin, catechin, rutin, and genistein,

- the at least one polyphenol which is part of the extract of the seed of the fruit of Linum usitatissimum is a lignan, preferably selected from the group consisting of secoisolariciresinol diglucoside, matairesinoside, lariciresinol, pinoresinol, sesamin, and secoisolariciresinol,

- the at least one polyphenol which is part of the extract of the root of Polygonum cuspidatum preferably selected from the group consisting of resveratrol, piceatannol, pinosylvin, pterostilbene, and rhapontienin.

30. The composition or kit of parts according to any of claims 28 or 29, wherein:

- the extract of the skin of the fruit of Prunus dulcis contains between 5-60% w/w, preferably 30% w/w of flavonoids, wherein the flavonoids are preferably selected from the group consisting of procyanidins, propelargonidins, prodelphinidins, catechin, epicatechin, quercetin, kaempferol, isorhamnetin, rutin, genistein, and combinations thereof,

- the extract of the fruit of Citrus paradisi or the extract of the seed of the fruit of Citrus paradisi contains 10-80% w/w, preferably 45% (w/w) of flavonoids, wherein the flavonoids are preferably selected from the group consisting of naringin, naringenin, quercetin, catechin, rutin, genistein, and combinations thereof,

- the extract of the seed of the fruit of Linum usitatissimum containing between 5-50% w/w, preferably 20% (w/w) of lignans, wherein the lignans are preferably selected from the group consisting of secoisolariciresinol diglucoside, matairesinoside, lariciresinol, pinoresinol, sesamin, and secoisolariciresinol,

- the extract of the root of Polygonum cuspidatum contains between 50-100% w/w, preferably 98% (w/w) of stilbenes, wherein the stilbenes are preferably selected from the group consisting of resveratrol, piceatannol, pinosylvin, pterostilbene, rhapontienin, and combinations thereof.

31. The composition or kit of parts according to any of claims 18-30, wherein the hydroxytyrosol content is at least 0.05-10% w/w, preferably at least 0.9% w/w with respect to the total of the composition or of the components of the kit of parts, and wherein the content in at least one polyphenol other than hydroxytyrosol is at least 1- 95% w/w, preferably at least 26% w/w with respect to the total of the composition or of the components of the kit of parts.

32. The composition or kit of parts according to any of claims 18-31, wherein the hydroxytyrosokpolyphenol w/w ratio is between 1:1 and 1:50, preferably 1:24.3, more preferably 1:28.

33. The composition according to any of claims 18-32, wherein the content of hydroxytyrosol in the composition is at least 0.05-10% w/w, preferably at least 0.9% w/w with respect to the total of the composition, wherein at least one polyphenol other than hydroxytyrosol is a polyphenol composition of almond skin, wherein the content of the polyphenol composition of almond skin is at least 10-50% w/w, preferably at least 26% w/w with respect to the total of the composition.

34. A method for obtaining a composition according to any of claims 18 to 33, which method comprises contacting a composition comprising hydroxytyrosol with a composition comprising at least one polyphenol that is not hydroxytyrosol and that does not occur naturally in the tree O. europaea.

35. The method according to claim 34 wherein the composition comprising hydroxytyrosol is a plant extract and/or wherein the composition comprising the at least one polyphenol is a plant extract.

36. The method according to claim 35 wherein the plant extract comprising hydroxytyrosol is an extract of the leaf or of the fruit of the tree O. europaea , and preferably contains 1-90% w/w, preferably 10% (w/w) of this compound.

37. The method according to claims 34 to 36, wherein the plant extract comprising at least one polyphenol is selected from the group consisting of:

- an extract of the skin of the fruit of Prunus dulcis

- an extract of the fruit of Citrus paradisi or the extract of the seed of the fruit of Citrus paradisi

- an extract of the seed of the fruit of Linum usitatissimum

- an extract of the root of Polygonum cuspidatum.

38. The method according to claim 37, wherein:

- the extract of the seed of the fruit of Linum usitatissimum containing between 5-50% w/w, preferably 20% (w/w) of lignans, wherein the lignans are preferably selected from the group consisting of secoisolariciresinol diglucoside, secoisolariciresinol, matairesinoside, lariciresinol, pinoresinol, and sesamin,

- the extract of the root of Polygonum cuspidatum contains between 50-100% w/w, preferably a 98% (w/w) of stilbenes, wherein the stilbenes are preferably selected from the group consisting of resveratrol, piceatannol, pinosylvin, pterostilbene, rhapontienin, and combinations thereof.

39. A pharmaceutical composition comprising a composition according to any of claims 18 to 33, or a composition obtained by the method according to claims 34 to 38, and a pharmaceutically acceptable excipient.

40. A food or nutritional supplement comprising a composition according to any of claims 18 to 33, or a composition obtained by the method according to claims 34 to 38 and a nutritionally acceptable excipient.

41. The composition or kit of parts according to any of claims 18 to 33, the composition obtained by the method according to claims 34 to 38, the pharmaceutical composition according to claim 39, or the food or nutritional supplement according to claim 40 for use in medicine.

42. The composition or kit of parts according to any of claims 18 to 33, the composition obtained by the method according to claims 34 to 38, the pharmaceutical composition according to claim 39, or the food or nutritional supplement according to claim 40 for use in the prevention and/or treatment of a cardiovascular disease.

43. The composition, kit of parts, the pharmaceutical composition, or the food or nutritional supplement for use according to claim 42, wherein the cardiovascular disease is selected from the group consisting of atherosclerosis, cerebrovascular accident, peripheral vascular disease, and coronary disease.

44. The composition, kit of parts, the pharmaceutical composition, or the food or nutritional supplement for use according to any of claims 41-43, wherein the composition, the kit of parts, the pharmaceutical composition or the food or nutritional supplement is administered to a patient with hypercholesterolemia, preferably with moderate hypercholesterolemia.

45. The kit of parts for use according to any of claims 41-44, wherein the hydroxytyrosol and the at least one polyphenol are administered separately.

46. The composition, the kit of parts, the pharmaceutical product, or the food or nutritional supplement for use according to any of claims 41-44, wherein the administration of the composition, of the components comprised in the kit of parts, of the pharmaceutical product, or of the food or nutritional supplement comprises administering 7.5 mg/day of hydroxytyrosol and 210 mg/day of polyphenols other than hydroxytyrosol.

47. The composition, the kit of parts, the pharmaceutical product, or the food or nutritional supplement for use according to any of claims 41-44, wherein the administration of the composition, of the components comprised in the kit of parts, of the pharmaceutical product, or of the food or nutritional supplement comprises administering 67 mg/day of plant extract providing hydroxytyrosol and 700 mg/day of plant extract providing polyphenols other than hydroxytyrosol.

48. The composition, the kit of parts, the pharmaceutical product, or the food or nutritional supplement for use according to any of claims 41-47, wherein the administration of the composition, of the components comprised in the kit of parts, of the pharmaceutical product, or of the food or nutritional supplement comprises administering multiple doses of the compositions, of the components comprised in the kit of parts, of the pharmaceutical product, or of the food or nutritional supplement for at least 1 week.

49. The composition, the kit of parts, the pharmaceutical product, or the food or nutritional supplement for use according to any of claims 41-48, wherein the administration of the composition, of the components comprised in the kit of parts, of the pharmaceutical product, or of the food or nutritional supplement comprises administering at least one daily dose, preferably 2 daily doses, of the composition, of the components comprised in the kit of parts, of the pharmaceutical product, or of the food or nutritional supplement.

50. The kit of parts for use according to any of claims 41-49 wherein the components of the kit are combined before the administration.