BE1031391B1 - COMPOSITION AND ORAL DOSAGE FORM - Google Patents

Abstract

The present invention relates to a composition comprising a mesoporous silica having a specific surface area, calculated from nitrogen sorption isotherms at liquid nitrogen temperature (77K) according to the Brunauer, Emmett and Teller (BET) theory, of at least 500 m²/g, wherein said mesoporous silica is impregnated with phloretin, glycosylated phloretin or a combination thereof, said glycosylated phloretin being phlorizin and phloretin 3′,5′-di-C-glucoside. The invention further relates to an oral dosage form, the use of said composition and a method for producing said composition.

Description

1 BE2023/5366

COMPOSITION AND ORAL DOSAGE FORM

FIELD OF THE INVENTION

The present invention relates to a composition and an oral dosage form. In another aspect, the present invention also relates to a process for preparing said composition and oral dosage form.

CONTEXT

Phloretin is a dihydrochalcone and is present in commonly consumed fruits such as apples and strawberries. Phloretin is known for its immunomodulatory properties and is widely used as a beauty product due to its antioxidant properties. Furthermore, phloretin has been shown, for example in WO2022157381, to play a role in CNS repair, including promoting and accelerating remyelination.

The improvement in repair was attributed to the ability of phloretin to stimulate OPC maturation using in vitro OPC cultures and ex vivo brain slices devoid of microglia.

However, although phloretin is a natural compound, its presence in plant derivatives is low, and it is absorbed slowly and eliminated quickly.

Thus, to obtain the positive health effects of phloretin, it is usually necessary to consume it long-term and/or at high levels.

The inclusion of phloretin and its derivatives in nutritional and pharmacological products further raises the problem of its low water solubility. The low water solubility is accompanied by low bioavailability of phloretin and its derivatives when administered orally, particularly in solid form. WO2022157381 does not address this issue.

There is a need for new techniques and products to deliver phloretin to animal and human populations and to improve the bioavailability of phloretin to animals and humans.

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SUMMARY OF THE INVENTION

The present invention and its embodiments provide a solution to one or more of the above-mentioned drawbacks. To this end, the present invention relates to a composition according to claim 1. The composition comprises a mesoporous silica impregnated with phloretin and/or a phloretin derivative. Preferred embodiments of the composition are described in any one of claims 2 to 7.

The composition according to claim 1 provides a significantly increased dissolution rate of phloretin and/or its derivatives compared to their unmodified counterparts. In addition, the total amount of phloretin dissolved is higher.

In a second aspect, the present invention relates to an oral dosage form according to claim 8. A preferred embodiment of the oral dosage form is set forth in claim 9. The significant increase in dissolution rate as well as increased amount of dissolved phloretin results in higher bioabsorption, thereby improving absorption in the gastrointestinal tract.

In a third aspect, the invention relates to a composition for use in the treatment and/or prevention of a demyelinating disease according to claim 10. Preferred embodiments of the composition for use are illustrated in any one of claims 11 to 14. In a fourth aspect, the invention relates to a method for producing a composition according to claim 15. Preferred embodiments of the method are described in claims 16 and 17. The improvement in absorption kinetics results in a greater efficiency of the use of phloretin and its derivatives in medical treatments. Furthermore, the dosage response is more stable. Without being bound by theory, it is believed that by radically improving bioabsorption, the absorbed phloretin more closely matches the ingested phloretin, rather than being bound by kinetics that may vary from case to case. In particular, the effects of natural differences in absorption due to, for example, the time of ingestion (especially in relation to meals) and physical differences are reduced.

This allows for more precise dosing as well as potentially higher dosing without causing side effects.

; ; 3 BE2023/5366

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to a composition comprising a mesoporous silica impregnated with phloretin and/or a phloretin derivative.

Unless otherwise defined, all terms used in the disclosure of the invention, including technical and scientific terms, have the meaning commonly understood by a person of ordinary skill in the art to which the present invention pertains. For the purpose of further guidance, definitions of terms are included to better appreciate the teaching of the present invention.

In this document, the following terms have the following meanings:

The term "phloretin" as used herein means the dihydrochalcone having the CAS number 60-82-2 and the IUPAC name 3-(4-hydroxyphenyl)-1-(2,4,6-trihydroxyphenyl)-propan-10ne.

A "glycoside" is a molecule in which a sugar group is linked to another functional group by a glycosidic bond. In formal terms, a glycoside is a — molecule in which a sugar group is linked by its anomeric carbon to another group via a glycosidic bond. Glycosides can be linked by an O- (an O-glycoside), N- (a glycosylamine), S- (a thioglycoside), or C- (a

C-glycoside). According to VIUPAC, the term “C-glycoside” is a misnomer; the preferred term is “C-glycosyl compound”. Preferably, the sugar is linked to a non-sugar so that the molecule is considered a glycoside, which excludes polysaccharides. The sugar group is then called a glycone and the non-sugar group is called an aglycone or genin portion of the glycoside. The glycone can consist of a single sugar group (monosaccharide) or multiple sugar groups (disaccharide or oligosaccharide).

The terms “glycosylated phloretin” and “phloretin glycoside,” as used herein, are synonymous and refer to a molecule in which one or more sugars are linked to one or more functional groups of a phloretin molecule via a glycosidic bond. The glycosides may be linked by an O- (an O-glycoside) or C- (a C-glycoside) glycosidic bond. The sugar is preferably a monosaccharide or a disaccharide. Preferably, the sugar(s) are glucoses.

4 BE2023/5366

The term "phlorizin" as used herein means a glucoside of phloretin having the CAS number 60-81-1 and the IUPAC name 1-(2,4-Dihydroxy-6-{[(2S,3R,45,5S,6R)-3,4,5-trihydroxy-6-(hydroxymethyl)oxan-2-yl]oxy}phenyl)-3-(4-hydroxyphenyl)propan-1-one).

The terms “a”, “an” and “the” and “the”, as used herein, refer to both the singular and the plural, unless the context clearly indicates otherwise. For example, “a compartment” means one or more compartments.

The term “about,” as used herein in reference to a measurable value such as a parameter, quantity, time duration, etc., is intended to encompass variations of +/-20% or less, preferably +/-10% or less, more preferably +/-5% or less, even more preferably +/-1% or less, and even more preferably +/-0.1% or less of the specified value, to the extent that such variations are relevant to the disclosed invention. However, it is understood that the value to which the modifier “about” refers is itself specifically disclosed.

The terms “include,” “comprising,” “comprises,” and “composed of” as used herein are synonymous with “comprising,” “comprising,” “comprises,” or “contain,” “contains,” and are inclusive or open terms that specify the presence of the following, e.g., components, and do not exclude or preclude the presence of additional components, features, elements, components, or steps not cited, known in the art, or disclosed herein.

Further, the terms first, second, third and the like in the description and claims are used to distinguish like elements and not necessarily to describe a sequential or chronological order, unless otherwise indicated. It is understood that the terms so used are interchangeable in appropriate circumstances and that the embodiments of the invention described herein may operate in sequences other than those described or illustrated herein.

Enumerating numeric ranges by their endpoints includes all numbers and fractions within that range, as well as the enumerated endpoints.

The term “wt%”, “weight percentage”, “wt%” or “wt%”, herein and throughout the specification, unless otherwise defined, refers to the relative weight of the respective component to the total weight of the formulation. 5

Although the terms "one or more" or "at least one", such as one or more elements or at least one element of a group of elements, are clear in themselves, in order to be even more precise, the term encompasses in particular a reference to any one of said elements, or to any two or more of said elements, such as, for example, 23, 24, 25, 26 or >7, etc. of such elements, and up to all of such elements

Unless otherwise defined, all terms used in the disclosure of the invention, including technical and scientific terms, have the meaning commonly understood by a person of ordinary skill in the art to which the present invention pertains. To better understand the teaching of the present invention, definitions of the terms used in the description are included for convenience.

The terms or definitions used herein are intended only to facilitate the understanding of the invention.

Reference throughout this specification to "an embodiment" means that a particular element, structure, or feature described in connection with the embodiment is included in at least one embodiment of the present invention. Thus, the presence of the phrase "in an embodiment" at various locations in this specification does not necessarily always refer to the same embodiment, although it may. Furthermore, the elements, structures, or features may be combined in any suitable manner, as would be apparent to a person skilled in the art from the present disclosure, in one or more embodiments. Furthermore, while some embodiments described herein include certain elements, but not other elements included in other embodiments, combinations of elements from different embodiments are intended to fall within the scope of the invention and form different embodiments, as would be understood by those skilled in the art. For example, in the following claims, any of the claimed embodiments may be used in any combination.

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In a first aspect, the invention relates to a composition.

In a preferred embodiment of the invention, said composition comprises a mesoporous silica impregnated with phloretin, a phloretin derivative or a combination thereof.

The phloretin derivative is preferably a glycosylated phloretin. In another preferred embodiment, the glycosylated phloretin is selected from the following list: phlorizin, phloretin 3',5'-di-C-glucoside and naringin dihydrochalcone, preferably the glycosylated phloretin is phlorizin.

In a particularly preferred embodiment of the invention, said composition comprises a mesoporous silica impregnated with phloretin, a glycosylated phloretin or a combination thereof. In an even more preferred embodiment of the invention, said composition comprises a mesoporous silica impregnated with phloretin, phlorizin or a combination thereof, preferably said mesoporous silica is impregnated with phloretin.

The mesoporous silica of the invention preferably has a specific surface area, calculated from nitrogen sorption isotherms at liquid nitrogen temperature (77K) according to the Brunauer, Emmett and Teller (BET) theory, of at least 10 m2/g, preferably at least 100 m2/g, more preferably at least 250 m2/g, even more preferably at least 500 m2/g. The specific surface area is preferably at most 1500 m2/g, more preferably at most 1250 m2/g, even more preferably at most 1000 m2/g, and even more preferably at most 800 m2/g.

In another or further embodiment of the invention, said mesoporous silica has a specific surface area, calculated from the — nitrogen sorption isotherms at liquid nitrogen temperature (77K) according to the Brunauer, Emmett and Teller (BET) theory, of between 10 and 1500 m2/g, preferably between 100 and 1250 m2/g, more preferably between 250 and 1000 m2/g, even more preferably between 500 and 800 m?/g.

The inventors found that this specific surface area allows for optimal release of phloretin, phloretin derivative or the combination thereof when dissolved in water.

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In a particularly preferred embodiment, the present invention provides a composition comprising a mesoporous silica having a specific surface area, calculated from nitrogen sorption isotherms at liquid nitrogen temperature (77K) according to the Brunauer, Emmett and Teller (BET) theory, of at least 500 m2/g, wherein said mesoporous silica is impregnated with phloretin, a phloretin derivative or a combination thereof.

The inventors found that gradual dissolution of a mesoporous silica impregnated with phloretin, a phloretin derivative or a combination thereof allowed for a gradual sustained release of said phloretin, a phloretin derivative or a combination thereof. This was found to improve the dissolution of said phloretin, a phloretin derivative or a combination thereof in aqueous environments, thereby improving their bioavailability.

In a preferred embodiment, the present invention provides an oral dosage form wherein said mesoporous silica has mesopores with an average pore size of at least 1 nm. The pore size was calculated from the nitrogen sorption isotherms at liquid nitrogen temperature (77K) according to the method of Barrett, Joyner and Halenda (BJH). The mesopore size of the silica, established from the nitrogen sorption isotherm at 77K, is 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25 nm or any value between the above values.

In another preferred embodiment, said mesoporous silica has mesopores whose average size, calculated from the nitrogen sorption isotherms at the temperature of liquid nitrogen (77K) according to the Barrett, Joyner and Halenda (BJH) method, is between 1 and 6 nm, preferably between 1 and 3 nm.

The inventors found that gradual dissolution of a mesoporous silica—impregnated with phloretin, a phloretin derivative, or a combination thereof—enables gradual sustained release of said phloretin, a phloretin derivative, or a combination thereof. This was found to enhance the dissolution of said phloretin, a phloretin derivative, or a combination thereof in aqueous environments, thereby improving their bioavailability.

In a preferred embodiment, the present invention provides a composition according to the first aspect of the invention, whereby said mesoporous silica has a pore volume of at least 0.1 cm3/g. In a preferred embodiment, the

8 BE2023/5366 pore volume of silica was calculated from nitrogen sorption isotherms at liquid nitrogen temperature (77K) by the method of Barrett, Joyner and Halenda (BJH). Said pore volume is at least 0.1 cm3/g, 0.2 cm3/9, 0.4 cm3/g, 0.5 cm3/g and at most 0.6 cm3/g, 0.8 cm3/g, 1.0 cm3/g, 1.2 cm3/9, 1.5 cm3/g or any value between the above values.

Phloretin, phloretin derivative, or combination thereof may be impregnated onto and/or into silica in a variety of ways. For example, the ingredient(s) may be deposited on the surface of the silica particles, incorporated into the pores of the porous silica, incorporated into pores formed by agglomeration or consolidation of the silica particles (particularly nanoparticles) or otherwise bound or associated with the surface of the silica. Suitable methods for loading the ingredient with the silica include drum coating, fluidized bed, spray drying, spray cooling, coating, dusting/breading, coextrusion techniques. Of these physical methods, spray drying of a silica suspension is preferred. These are well known techniques, details of which can be found in standard texts, for example, for a number of these techniques see Food Processing Technology, Principles and Practice by PJ Fellows, second edition, Woodhead Publishing Ltd.

In a preferred embodiment, the phloretin, phloretin derivative or combination thereof may be impregnated onto the mesoporous silica as follows: the phloretin, phloretin derivative or combination thereof to be loaded onto the silica may be dissolved or suspended in a suitable solvent, and the silica particles may be incubated in the resulting solution for a suitable period of time. Aqueous and non-aqueous slurries have been produced from ground silica powder and the processing and properties of the silica suspensions have been studied and reported by Sacks in Ceram. Eng. Sci. Proc. 6, 1985, pp. 1109-23 and Kerkar in J. Am. Chem. Soc. 73, 1990, pp. 2879-85. Removal of the solvent results in deposition of the ingredient on the surface of the silica particles. In the case of mesoporous silica, the ingredient solution penetrates into the pores of the silica by capillarity and, after removal of the solvent, the ingredient is present in the pores. The preferred solvents are water, ethanol and isopropyl alcohol, GRAS solvents and volatile liquids which are suitable for lyophilization. It is clear that the solvent used must be sufficiently miscible with the phloretin, the phloretin derivative or their combination to be impregnated on the silica.

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Higher loading levels, e.g. at least 15% by weight of

The bulk charge relative to the weight of the silica charged, can be obtained by carrying out the impregnation at an elevated temperature. For example, the charge can be carried out at a temperature equal to or higher than the melting point of the ingredient to be charged. Quantification of the bulk charge can be conveniently achieved by a number of known analytical methods, including gravimetry, VEDX (energy dispersive X-ray analysis),

Fourier transform infrared (FTIR), Raman spectroscopy, UV spectrophotometry, titrimetric analysis, HPLC or mass spectrometry. — If necessary, quantification of charge uniformity can be achieved by techniques capable of spatial resolution, such as cross-sectional EDX, Auger depth profiling, micro-Raman and micro-FTIR.

Loading levels can be determined by dividing the weight of the ingredient — absorbed during loading (equivalent to the mass of the ingredient absorbed divided by its density) by the void volume of the porous silica before loading, multiplied by one hundred.

In one embodiment, the mesoporous silica is subjected to a heat treatment between 450°C and 950°C. More preferably, said mesoporous silica is subjected to a heat treatment between 450°C and 700°C, and even more preferably between 500°C and 600°C. Most preferably, said mesoporous silica is subjected to a heat treatment at 500°C, 510°C, 520°C, 530°C, 540°C, 550°C, 560°C, 570°C, 580°C, 590°C or 600°C.

In one embodiment of the invention, the mesoporous silica is a plugged mesoporous silica. The term "plugged mesoporous silica" as used herein refers to a mesoporous silica comprising a surfactant in at least some of its mesopores. A "plugged mesoporous silica" is not limited to a mesoporous silica in which all or substantially all of the mesopores are plugged. Without being bound by theory, it is believed that the surfactant contained in the mesoporous silica partially plugs these mesopores. In other words, the mesopores are at least partially blocked or otherwise unavailable.

It should be noted that this "clogging" does not require that all pores, or even all mesopores, be unavailable. Furthermore, the pores do not need to be permanently clogged. In fact, the clogging is supposed to be reversible, that is, the mesopores can be unclogged. For example, the mesopores

10 BE2023/5366 can be at least partially unclogged for example by washing away said surfactant.

In another instance, the surfactant is an amphiphilic glycoside. In another preferred embodiment, the surfactant is an amphiphilic glycoside combined with fatty acid esters. Amphiphilic glycosides, alone or in combination with fatty acid esters (such as commercial "Tween"), have been found to be particularly preferable to other surfactants because this class of compounds are quite effective at calcining at the temperatures indicated above.

An amphiphilic compound is a chemical compound that has both hydrophilic and hydrophobic or lipophilic properties. Such a compound is called amphiphilic or amphipathic. This forms the basis of a number of research areas in chemistry and biochemistry. Common amphiphilic substances are also called surfactants or substances, soaps, detergents, and lipoproteins.

Preferred amphiphilic compounds include many biological compounds such as, but not limited to: phospholipids, cholesterol, glycolipids, fatty acids, saponins, etc. Particularly preferred amphiphilic compounds are glycosides.

In a more preferred embodiment, the present invention provides a composition, wherein said amphiphilic glycoside is saponin. Saponins are a class of chemical compounds found in abundance in various plant species. More specifically, they are amphipathic glycosides grouped phenomenologically by the soapy foam they produce when agitated in aqueous solutions, and structurally by the association of one or more hydrophilic glycoside moieties with a lipophilic triterpene derivative.

The main sources of saponins used in the food and cosmetic industry are the tree Quillaja saponaria Molina, Yucca schidigera, the Southeast Asian shrub Camellia sinensis, known as the tea plant, and the leaves of Hedera helix L., known as ivy leaves. The inventors have discovered that the cooperative self-assembly mechanism can be achieved in the presence of natural saponin-type surfactants. It appears that, as in the

11 BE2023/5366 case of “classic” industrial surfactants, the saponins aggregate to form, with the silica precursor, a hybrid phase on which the synthesis of the mesoporous material will take place.

The saponins used in the context of the present invention are triterpenoid glycosides whose main structure (I) aglycone (sapogenin) can take the following forms: dammarenediols (dammaranes), cucurbitadienol (cucurbitanes), hopanol (hopanes), lanosterols (lanostanes), tirucalladienol (tirucallanes), B-amyrin (oleananes), oo-amyrin (ursanes), taraxasterols — (taraxasterols), lupeol (lupans).

Methyl radicals, carboxylic functions, an aldehyde or an alcohol; hydrogen atoms, hydroxyl groups, as well as simple or branched osidic chains are also attached to the aglycone skeleton. Within the osidic chains, the following sugars are favored: D-glucose, L-rhamnose, D-galactose, D-glucuronic acid, L-arabinose, D-xylose, D-fructose, D-adipose, D-fucose. In the case where the aglycone contains an osidic chain, it is called a monodesmoside and in the case where two glycoside chains are linked to the aglycone skeleton, it is called a bidesmoside.

In a preferred embodiment, the present invention provides a composition comprising said mesoporous silica in an amount of at least 1% by weight, relative to the total weight of said composition, preferably at least 25% by weight, more preferably at least 50% by weight, even more preferably at least 60% by weight, even more preferably at least 65% by weight.

In another or additional embodiment, said composition comprises said mesoporous silica in an amount of at most 99% by weight relative to the total weight of said composition, preferably at most 90% by weight, more preferably at most 85% by weight, even more preferably at least 80% by weight, even more preferably at most 75% by weight.

In another or additional embodiment, said composition comprises said mesoporous silica in an amount of between 1 and 99% by weight, relative to the total weight of said composition, preferably between 25 and 90% by weight, more preferably between 50 and 85% by weight, still

12 BE2023/5366 more preferably between 60 and 80% by weight, even more preferably between 65 and 75% by weight.

In a preferred embodiment, the present invention provides a composition - comprising said phloretin, said phloretin derivative or said combination thereof in an amount of at least 1% by weight, relative to the total weight of said composition, preferably at least 10% by weight, more preferably at least 20% by weight.

In another preferred embodiment or a further preferred embodiment, said composition comprises said phloretin, said phloretin derivative or said combination thereof in an amount of at most 99% by weight relative to the total weight of said composition, preferably at least 50% by weight, more preferably at least 40% by weight.

In another preferred embodiment or a further preferred embodiment, said composition comprises said phloretin, said phloretin derivative or said combination thereof in an amount of between 1 and 99% by weight, relative to the total weight of said composition, preferably between 10 and 50% by weight, more preferably between 20 and 40% by weight.

In a preferred embodiment of the invention, the weight ratio of the total amount of said phloretin and said phloretin derivative to said mesoporous silica is at least 1/99, preferably at least 10/90, more preferably at least 20/80, even more preferably at least 25/75.

In another preferred embodiment or a further preferred embodiment of the invention, the weight ratio of the total amount of said phloretin and said phloretin derivative to said mesoporous silica is at most 60/40, preferably at most 50/50, more preferably at least 40/60, even more preferably at least 35/65.

In another preferred embodiment or a further preferred embodiment of the invention, the weight ratio of the total amount of said phloretin and said phloretin derivative to said mesoporous silica is between 1/99 and 60/40, preferably between 10/90 and 50/50, more preferably between 20/80 and 40/60, and even more preferably between 25/75 and 35/65.

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In a preferred embodiment of the invention, at least 60% by weight of said phloretin or said phloretin derivative is dissolved in water after 2 hours at 37°C. This means that when the composition of the present invention is introduced into water at 37°C, at least 60% by weight of the phloretin and/or the phloretin derivative is dissolved in the water after 2 hours. In another preferred embodiment, at least 65% by weight, more preferably at least 70% by weight, more preferably at least 75% by weight, more preferably at least 80% by weight, more preferably at least 85% by weight, more preferably at least 90% by weight, more preferably at least 95% by weight of said phloretin or said phloretin derivative is dissolved in water after 2 hours at 37°C. This high amount of dissolved phloretin or phloretin derivative results in higher bioavailability.

In a preferred embodiment of the invention, the phloretin or phloretin derivative which is impregnated in or on the mesoporous silica has an XRD crystallinity of at most 80%, preferably at most 70%, more preferably at most 60%, more preferably at most 50%, more preferably at most 40%, more preferably at most 35%, more preferably at most 30%, more preferably at most 25%, more preferably at most 20%, more preferably at most 15%, more preferably at most 10%, more preferably at most 8%, more preferably at most 7%, more preferably at most 6%, most preferably at most 5%.

The degree of XRD crystallinity of the samples can be determined by the X-ray diffraction method described below. The inventors found that lower XRD crystallinity was associated with a significant increase in the dissolution rate.

In a preferred embodiment of the invention, the phloretin or phloretin derivative has a median particle size (d50), as determined by SEM, of the phloretin or glycosylated phloretin of at most 30 µm, more preferably at most 25 µm, even more preferably at most 20 µm, even more preferably at most 15 µm, even more preferably at most 10 µm, even more preferably at most 5 µm, most preferably at most 1 µm.

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In a preferred embodiment of the invention, the phloretin or phloretin derivative has a particle size d90, as determined by SEM, of the phloretin or glycosylated phloretin of at most 30 µm, more preferably at most 20 µm, even more preferably at most 10 µm, even more preferably at most 1 µm.

Smaller particle sizes result in significantly higher dissolution rates. Without being bound by theory, it is assumed that this effect may be caused by the significant increase in surface area, a decrease in surface tension, an associated decrease in crystal formation and lattice energy.

In a particularly preferred embodiment, the present invention provides a composition consisting of a mesoporous silica having a specific surface area, — calculated from nitrogen sorption isotherms at liquid nitrogen temperature (77K) according to the Brunauer, Emmett and Teller (BET) theory, of at least 500 m2/g, wherein said mesoporous silica is impregnated with phloretin, a phloretin derivative or a combination thereof. In a preferred embodiment, the surface area is at least 550 m2/g, more preferably at least 600 m2/g, more preferably at least 650 m2/g, more preferably at least 700 m2/g, more preferably at least 750 m2/g, more preferably at least 800 m2/g, more preferably at least 850 m2/g, more preferably at least 900 m2/g, more preferably at least 950 m2/g, more preferably at least 1000 m2/g. The applicant has found that an increase in the surface area of the mesoporous silica increases the dissolution rate as well as the dissolution equilibrium of the phloretin dispersed thereon.

In another particularly preferred embodiment, the present invention—provides a composition comprising a mesoporous silica having a specific surface area, calculated from nitrogen sorption isotherms at liquid nitrogen temperature (77K) according to the Brunauer, Emmett and Teller (BET) theory, of at least 500 m2/g, wherein said mesoporous silica is impregnated with phloretin, a phloretin derivative or a combination thereof, the composition comprising said mesoporous silica in an amount of from 60 to 80% by weight, and said phloretin, said phloretin derivative or said combination thereof in an amount of from 20 to 40% by weight.

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Phloretin can be obtained and purified from nature or synthesized. In a preferred embodiment, highly pure extracts can be produced from natural extracts comprising phloretin. More preferably, said natural extracts comprising phloretin are derived from apple trees (Malus, preferably Malus Pumila), strawberry trees (Fragaria) or apricot trees (Prunus, preferably Prunus mandshurica). In another preferred embodiment, the natural extract comprising phloretin is produced from the leaves, bark or root of a fruit plant, preferably the aforementioned fruit plants. Most preferably, the root of a fruit plant, in particular the root of Malus pumila. The root of Malus pumila contains a relatively high concentration of phloretin. More importantly, it can be efficiently purified to obtain a highly pure phloretin extract.

Advantageously, naturally occurring phloretin is ecologically beneficial.

Naturally occurring phloretin extracts typically include a mixture of phloretin and glycosylated phloretin. Glycosylated phloretin can be converted to phloretin by hydrolysis, such as enzymatic hydrolysis or

Acid hydrolysis. Enzymatic hydrolysis offers ecological advantages.

Acid hydrolysis can achieve better conversion and higher product purity.

In a more preferred embodiment, the composition further comprises a salt, said salt consisting of a cation and an anion, said salt being selected from the list of alkali metal salts, alkaline earth metal salts or a mixture thereof. More preferably, the salt is selected from magnesium salts, calcium salts, potassium salts, sodium salts or a mixture thereof, more preferably magnesium salts or calcium salts, most preferably calcium salts. Preferably, said salt is an organic salt, preferably said salt is an acetate, a citrate, a tartrate, a formate, a benzoate, a gluconate, a sorbate or a mixture thereof. Most preferably, said salt is calcium acetate. Preferably, said salt is edible, more preferably said salt is pharmaceutically acceptable.

Preferably, the weight ratio between the cation of said salt and the mesoporous silica is between 1:100 and 200:100, preferably the weight ratio between the cation of said salt and the mesoporous silica is between 10:100 and 200:100, more preferably between 50:100 and 150:100. The inventors have surprisingly discovered that the addition of these salts, probably due to the stabilizing effects of silicic acids such as orthosilicic acid associated with the dissolved cations once in an aqueous medium, considerably increases the rate of

16 BE2023/5366 dissolution of mesoporous silica. This advantageously enables the composition to provide bioavailable phloretin or derivatives thereof as well as bioavailable silicon. Furthermore, the dissolution of the mesoporous silica facilitates the dissolution of the phloretin particles.

In a preferred embodiment, the composition further comprises piperine. The inventors have discovered that piperine acts as a synergist of phloretin and its derivatives. Without being bound by theory, it is believed that this effect is due to an increase in the absorption of phloretin and/or a decrease in the in vivo degradation of phloretin and its derivatives. In another preferred embodiment, the composition comprises piperine in an amount of 5 to 15% by weight. In a preferred embodiment, the mesoporous silica is impregnated with phloretin and piperine by grinding or solvent impregnation.

Preferably, the mesoporous silica is impregnated with phloretin and piperine by — milling, preferably by ball milling. In another embodiment, the piperine may be added by the addition of peppers. Suitable peppers include, but are not limited to, black pepper, white pepper, and long pepper.

In a preferred embodiment, the composition further comprises resveratrol, preferably trans-resveratrol. The inventors have found that resveratrol provides a series of nutritional and medical effects that synergize with resveratrol. In another preferred embodiment, the composition comprises resveratrol in an amount of 5 to 30% by weight, preferably in an amount of 5 to 15% by weight. In a preferred embodiment, the mesoporous silica is impregnated with phloretin and resveratrol by milling or solvent impregnation. Most preferably, the mesoporous silica is impregnated with both phloretin and resveratrol by milling, preferably ball milling. The result is a dispersion of resveratrol and phloretin, wherein the resveratrol is preferably also a fine particle dispersion. More preferably, the d50 of the resveratrol particles, as determined by SEM, is at most 30 um, more preferably at most 25 um, even more preferably at most 20 um, even more preferably at most 15 um, even more preferably at most 10 um, even more preferably at most 5 um, most preferably at most 1 um. — Advantageously, the phloretin, resveratrol and mesoporous silica can be impregnated in a single process. The bioavailability of both phloretin and resveratrol is significantly increased. The inventors have found that the combination of phloretin and resveratrol is particularly suitable for

17 BE2023/5366 treatment and prevention of cardiovascular diseases. In a particular embodiment, the invention relates to a composition comprising mesoporous silica, resveratrol and phloretin or an oral dosage form comprising this composition for medical use; preferably for the treatment or prevention of cardiovascular diseases.

In a second aspect, the present invention provides an oral dosage form comprising a composition according to the first aspect.

In a preferred embodiment, said oral dosage form is a solid oral dosage form such as a tablet or a powder, and more preferably, said solid oral dosage form is a tablet. For reasons of ease of dissolution, said tablet is preferably not provided with an additional coating.

The inventors have found that the composition according to the first aspect is particularly advantageous as an oral dosage form, because the inventors have found that the composition according to the first aspect enhances the absorption of phloretin, phloretin derivatives or combinations thereof. These effects are particularly pronounced in the gastrointestinal tract.

In a preferred embodiment, the present invention provides an oral dosage form comprising said composition in an amount of at least 1% by weight, relative to the total weight of said oral dosage form, preferably at least 25% by weight, preferably at least 50% by weight, preferably at least 80% by weight.

A higher percentage of the composition according to the invention allows a higher concentration of silicon and the drug to be delivered, i.e. — phloretin, phloretin derivative or their combination, in the gastrointestinal tract of the subject, and therefore allows better absorption in the human or animal body.

The oral dosage form according to the present invention may further comprise one or more of the following ingredients: oxygen-sensitive edible oils; minerals; oxygen-sensitive fats, including dairy fats; oil-soluble ingredients; vitamins; enzymes; probiotic bacteria; prebiotics; nutraceuticals; amino acids; herbal extracts; herbs;

18 BE2023/5366 plant extracts; edible acids; salt; antioxidants; and pharmaceutically active compounds.

Oxygen-sensitive edible oils

Oxygen-sensitive edible oils include polyunsaturated fatty acids, including canola oil, borage oil, evening primrose oil, safflower oil, sunflower oil, pumpkin seed oil, rosemary oil, rice bran oil, flaxseed oil, wheat germ oil, and grapeseed oil. Some of these oils contain linoleic acid, alpha-linolenic acid, oleic acid, palmitic acid, and stearic acid.

Also included are marine oils, for example those derived from fish such as tuna, herring, mackerel, sardines, cod liver and shark.

Minerals and trace elements

Suitable minerals include: macrominerals including Ca, P, Mg, Na,

K; microminerals including Fe, Zn, Cu, Se, Cr, I, Mn, Mo, F. Suitable trace elements include Ni, V, B, Co.

Mixing and compressing certain metal powders with silicon powder can be used to create edible microbatteries. After ingestion, the gastrointestinal fluid acts as an electrolyte. If the metal is less noble than silicon, the resulting galvanic coupling increases the dissolution rate of this metal, which increases bioavailability.

Vitamins

Suitable vitamins include ascorbic acid, beta-carotene, biotin, choline, folic acid, niacin, pantothenic acid (vitamin B5), phylloquinone (vitamin K), pyridoxine (vitamin B6), riboflavin (vitamin B2), thiamine (vitamin B1), vitamin A, vitamin B12, vitamin D, vitamin

And mixtures thereof. The vitamin and silicon may be combined by allowing the vitamin to impregnate the silicon, optionally in the presence of gentle heat, typically between 40°C and 200°C.

Enzymes

Suitable enzymes are selected from the classes of carbohydrases, pectic enzymes, celluloses, proteases, oxidases and lipases. Examples include

Vamylase, bromelain, catalase, ficin, glucoamylase, glucose isomerase,

19 BE2023/5366 glucose oxidase, invertase, lactase, lipase, papain, pepsin, pullulanase and rennet.

Nutraceutical products

A nutraceutical ingredient provides medical or health benefits, including the prevention and treatment of disease. Typically, a nutraceutical is specifically tailored to provide the consumer with a particular health benefit. Nutraceuticals suitable for use in the present invention may be selected from curcumin (Curcuma longa), aloe vera (Aloe ferox, A. barbadensis), artichoke, Asian ginseng (Panax ginseng), astragalus, bee pollen, bilberry (Vaccinium myrtillus), black cohosh, cayenne pepper, hot pepper (Capsicum species), cascara sagrada (Rhamnus purshiana), cat's claw (Uncaria tomentosa), chamomile (Matricaria recutita), cranberry, dandelion (Taraxacum officinale), donq quai (Angelica sinensis), —echinacea (Echinacea purpurea and related species), evening primrose oil (Oenothera biennis), feverfew (Tanacetum parthenium), fructo-oligosaccharides, garlic (Allium sativum), ginger (Zingiber officinale), ginkgo (Ginkgo biloba), ginseng, glucarate, glucosamine, goldenseal (Hydrastis canadensis), gotu kola (Centella Asiatica), grape seed extract, green tea, guarana (Paullinacupana), hawthorn (Crataegus oxyacantha),

Vinositol, inulin, isoflavones, kava kava (Piper methysticum), L-carnitine, lecithin, licorice (Glycyrrhiza glabra and G. uralensis), lycopene, milk thistle (Silybum marianum), citrus peel mod., nettle, oligofructose,

Vomega-3, passion flower, Passion flower (Passiflora incarnata), pau d'arco, (Tabebuia impetiginosa), peppermint (Mentha piperita), phospholipids, polyphenol, psyllium (Plantago ovata and P. Major), pycnogenol, queroetin, D-limonene; reishi, ribonucleic acid, royal jelly, St. John's wort (Hypericum perforatum), saw palmetto (Serenoa repens; Sabal serrulata), schisandra, soy isoflavones, valerian turmeric (Valeriana officinalis) and mixtures thereof.

Amino acids

Amino acids suitable for use in the present invention may be selected from alanine, arginine, aspartic acid, asparagine, carnitine, cysteine, glutamic acid, glutamine, glutathione, glycine, histidine, isoleucine, leucine, lysine, methionine, ornithine, phenylalanine, proline, serine, taurine, threonine, tryptophan, tyrosine, valine and mixtures thereof.

20 BE2023/5366

Plant and herbal extracts

Suitable plant extracts include one or more plant sterols, including beta-sitosterol, campesterol, and stigmasterol. Suitable plant stanols include sitostanol, octacosanol, and policosanol. Suitable herbs include black walnut, burdock, chamomile, comfrey, echinacea, eucalyptus, hawthorn, hyssop, ginkgo, lemon balm, milk thistle, mullein, peppermint, psyllium, sage, saw palmetto, sheep sorrel, slippery elm, St. John's wort, thyme, turkey rhubarb, valerian, vitex.

Herbs suitable for medicinal use are described in The Natural Pharmacy by M. Polunin & C. Robbins (Dorling Kindersley 1999), 144 pages. In particular, pages 30 to 131 list suitable herbs. Suitable culinary herbs are described in Food Commodities, 2nd

Edition pp158-163 by B. Davis (Butterworth Heinemann 1994).

In a particular embodiment, the composition or oral dosage form of the present invention further comprises an essential oil derived from: Nigella sativa, Acorus gramineus, Lavandula angustifolia, Eucalyptus globulus, Mentha piperita, Rosmarinus officinalis, Jasminum sambac, Piper nigrum or a combination thereof. The addition of these essential oils is particularly advantageous for use in the treatment or prevention of demyelinating diseases. These essential oils have been found to have synergistic neuroprotective effects with phloretin.

Edible acids

Edible acids suitable for use in the present invention may be selected from citric acid, ascorbic acid, malonic acid, acetic acid, tartaric acid, sorbic acid, fumaric acid, malic acid, phosphoric acid, succinic acid and nicotinic acid.

Antioxidants

Antioxidants suitable for use in the present invention may be selected from sodium carbonate, calcium carbonate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithins, sodium lactate, calcium lactate, calcium malate and ammonium citrate.

21 BE2023/5366

The dihydrochalcone phloretin accelerates remyelination. In particular, phloretin stimulates OPC maturation in in vitro OPC cultures and enhances (re)myelination in ex vivo demyelinated brain slices devoid of microglia. Furthermore, phloretin was shown here to drive OPC differentiation via PPARγ activation.

Due to these effects on remyelination and oligodendrocyte differentiation and maturation, the present application provides, in a third aspect, a composition according to the first aspect or an oral dosage form according to the second aspect for medical use.

In a particularly preferred embodiment of the third aspect, the invention relates to a composition according to the first aspect or an oral dosage form according to the second aspect for use in the treatment and/or prevention of neurodegenerative diseases. These neurodegenerative diseases are generally characterized by a reduction or even a loss of function of the nerve cells of the brain or of the peripheral nervous system. Among the neurodegenerative diseases, mention may be made of Alzheimer's disease, dementia, multiple sclerosis, Parkinson's disease, ALS, Charcot-Marie disease,

Tooth, Huntington's disease, multiple system atrophy, traumatic nerve injury, diabetic neuropathy, chronic inflammatory polyradiculoneuropathy, Guillain-Barre syndrome.

In another embodiment, the present invention provides a composition according to the first aspect for use in the prevention and/or treatment of a neurodegenerative and demyelinating disease. The present invention therefore aims to provide a composition according to the first aspect and/or an oral dosage form according to the second aspect, preferably an oral dosage form according to the second aspect, for the prevention and/or treatment of a neurodegenerative and demyelinating disease of the central or peripheral nervous system.

In one embodiment, the neurodegenerative and demyelinating disease is a neurodegenerative and demyelinating disease of the central nervous system; preferably, said disease is multiple sclerosis; even more preferably, said disease is progressive multiple sclerosis. In another embodiment, the neurodegenerative and demyelinating disease is a neurodegenerative and demyelinating disease of the peripheral nervous system; in particular a neurodegenerative and demyelinating disease selected from diabetic neuropathy, Charcot-Marie Tooth disease or nerve injury.

22 BE2023/5366 traumatic, chronic inflammatory polyradiculoneuropathy, syndrome of

Guillain Barre.

As used herein, the term “demyelinating disease” refers to a pathological condition in which the myelin sheath surrounding neurons in nervous tissue is lost or damaged, resulting in axonal degeneration and impaired signal transduction in the affected nerves. A demyelinating disease of the central nervous system is a disease in which the myelin sheaths of neurons in the central nervous system are lost or damaged. Examples of demyelinating diseases of the central nervous system include multiple sclerosis, neuromyelitis optica (Devic's disease), inflammatory demyelinating diseases, central nervous system neuropathy, central pontine myelinolysis, myelopathy, leukoencephalopathy, or leukodystrophy.

A demyelinating disease of the peripheral nervous system is a pathological condition in which the myelin sheaths of neurons of the peripheral nervous system are lost or damaged. Demyelinating diseases of the peripheral nervous system are, for example, Guillain-Barre syndrome, chronic inflammatory demyelinating polyneuropathy, anti-MAG peripheral neuropathy, Charcot-Marie Tooth disease, hereditary neuropathy with hypersensitivity to pressure; conditions associated with copper deficiency, such as peripheral neuropathy, myelopathy and optic neuropathy; progressive inflammatory neuropathy, diabetic neuropathy or traumatic nerve injury. In another preferred embodiment of the third aspect, the invention relates to a composition according to the first aspect or an oral dosage form according to the second aspect for providing neuroprotective effects.

In this document, the term “multiple sclerosis” or “MS” refers to an autoimmune-mediated process in which an abnormal response of the body’s immune system is directed against the central nervous system (CNS), which consists of the brain, spinal cord, and optic nerves. The immune response results in the death of oligodendrocytes, demyelination, and eventual loss of axons, resulting in physical and cognitive disability.

In this document, the term “progressive multiple sclerosis” or “MS-

P » is characterized by an accumulation of chronic demyelinated lesions and is

23 BE2023/5366 subdivides into primary progressive MS (PPMS), secondary progressive MS (SPMS)

PS) and relapsing-remitting MS (RRMS).

Primary progressive MS (PPMS) is characterized by worsening of neurological function (accumulation of disability) from the onset of symptoms, without early relapses or remissions. PPMS can be further characterized at different time points as active (with occasional relapse and/or evidence of new MRI activity) or nonactive, as well as with progression (evidence of disease worsening as measured by an objective measure of change over time, with or without relapse or new MRI activity) or without progression. Secondary progressive MS (SPMS) follows an initial relapsing-remitting course. Most people diagnosed with relapsing-remitting MS (RRMS) eventually progress to a secondary progressive form in which there is progressive worsening of neurological function (accumulation of disability) over time. PS-MS can be further characterized at different time points as active (with relapses and/or evidence of new MRI activity) or nonactive, as well as with progression (evidence of disease worsening as measured by an objective measure of change over time, with or without relapses) or without progression.

The subject may be a non-human animal or a human being. Preferably, the subject is a human being.

In another preferred embodiment of the third aspect, the invention relates to a composition according to the first aspect or an oral dosage form according to the second aspect for use in the treatment and/or prevention of skin conditions. In particular those related to the health and well-being of the skin.

More particularly, for use in the treatment and prevention of skin aging, hyperpigmentation, acne and skin cancer.

In another preferred embodiment of the third aspect, the invention relates to a composition according to the first aspect or an oral dosage form according to the second aspect, for use in the treatment and/or prevention of liver diseases. In particular, the invention relates to a composition according to the first aspect or an oral dosage form according to the second aspect for hepatoprotective effects.

24 BE2023/5366

In another preferred embodiment of the third aspect, the invention relates to a composition according to the first aspect or an oral dosage form according to the second aspect, for use in the treatment and/or prevention of bone diseases. In particular, the invention relates to a composition according to the first aspect or an oral dosage form according to the second aspect, for use in the treatment or prevention of estrogen deficiency; more particularly for use in the treatment or prevention of osteoclastogenic resorption induced by estrogen deficiency.

In another preferred embodiment of the third aspect, the invention relates to a composition according to the first aspect or an oral dosage form according to the second aspect in the treatment and/or prevention of cardiovascular diseases. In particular, the invention relates to a composition according to the first aspect or an oral dosage form according to the second aspect for use in the treatment or prevention of cardiomyopathy.

In another preferred embodiment of the third aspect, the invention relates to a composition according to the first aspect or an oral dosage form according to the second aspect in the treatment and/or prevention of obesity. In particular, the invention relates to a composition according to the first aspect or an oral dosage form according to the second aspect for use in the treatment of adipose tissue. In a fourth aspect, the present invention provides a method for producing a composition according to the first aspect.

In a preferred embodiment of the invention, the method comprises the steps of: ii — providing a mesoporous silica, preferably with a specific surface area, calculated from nitrogen sorption isotherms at liquid nitrogen temperature (77K) according to the Brunauer, Emmett and Teller (BET) theory, of at least 500 m?/g; ii. impregnating said mesoporous silica with phloretin, a phloretin derivative or a combination thereof.

In another embodiment of the invention, said impregnation method is = selected from the following list: ball milling, hammer milling, twin screw granulation, certech milling, solvent impregnation, or a combination thereof, preferably ball milling, solvent impregnation or a combination thereof.

25 BE2023/5366

In a preferred embodiment of the invention, the method comprises the steps of: ii — providing a mesoporous silica, preferably with a specific surface area, calculated from nitrogen sorption isotherms at liquid nitrogen temperature (77K) according to the Brunauer, Emmett and Teller (BET) theory, of at least 500 m?/g; ii. impregnating said mesoporous silica with phloretin, a phloretin derivative or a combination thereof by ball milling.

In a preferred embodiment of the invention, the particle size d90, as measured by SEM, of the phloretin, phloretin derivative or combination thereof, is reduced by at least a factor of 2 during the impregnation in step ii, preferably by at least a factor of 10.

In a preferred embodiment of the invention, the XRD crystallinity, as determined using X-ray diffraction and Segal's formula, of phloretin, phloretin derivative or their combination is decreased by at least a factor of 1.25, preferably by at least a factor of 1.5, more preferably by at least a factor of 2.

In another preferred embodiment of the invention, the method comprises the steps of: ii. providing a mesoporous silica, preferably with a specific surface area, calculated from nitrogen sorption isotherms at liquid nitrogen temperature (77K) according to the Brunauer, Emmett and Teller (BET) theory, of at least 500 m?/g; ii. providing phloretin, a phloretin derivative or a combination thereof having a first particle size d90, as determined by SEM; iii. impregnating said mesoporous silica with phloretin, a phloretin derivative or a combination thereof, said phloretin, said phloretin derivative or said combination thereof having a second particle size d90, as determined by SEM, smaller than the first particle size d90.

In this embodiment of the invention, the ratio between said first particle size d90 and said second particle size d90 is at least 2, preferably at least 5, more preferably at least 10.

26 BE2023/5366

In another embodiment of the invention, the first particle size d90, as determined by SEM, of the phloretin or phloretin derivative is between 100 and 500 µm, preferably between 200 and 400, more preferably between 250 and 350 µm.

In another embodiment of the invention, the second particle size d90, as determined by SEM, of the phloretin or phloretin derivative is at most 30 um, more preferably at most 20 um, even more preferably at most 10 um, even more preferably at most 1 um.

The inventors found that by reducing the particle size of phloretin or phloretin derivative during impregnation into or onto mesoporous silica, the dissolution rate in water increased significantly. This higher dissolution rate results in higher bioavailability, and in particular, greater absorption of phloretin or phloretin derivative in the gastrointestinal tract.

In another preferred embodiment of the invention, the method comprises the steps of: ii. providing a mesoporous silica, preferably with a specific surface area, calculated from nitrogen sorption isotherms at liquid nitrogen temperature (77K) according to the Brunauer, Emmett and Teller (BET) theory, of at least 500 m?/g; ii. providing phloretin, a phloretin derivative or a combination thereof having a first XRD crystallinity, as determined using X-ray diffraction and the Segal formula; iii. — impregnating said mesoporous silica with phloretin, a phloretin derivative or a combination thereof, wherein said phloretin, said phloretin derivative or said combination thereof has a second XRD crystallinity, as determined using X-ray diffraction and the Segal formula, lower than the first XRD crystallinity.

In this embodiment of the invention, the ratio between the first crystallinity = XRD and the second crystallinity XRD is at least 1.25, preferably at least 1.5, more preferably at least 2.

27 BE2023/5366

In another embodiment of the invention, the first XRD crystallinity, as determined using X-ray diffraction and the Segal formula, of phloretin or phloretin derivative is between 70 and 100%, preferably between 80 and 100%, more preferably between 250 and 350 um.

In another embodiment of the invention, the second XRD crystallinity, as determined using X-ray diffraction and the Segal formula, of phloretin or phloretin derivative is at most 70%, preferably at most 60%, even more preferably at most 50%, even more preferably at most 40%.

The inventors discovered that by decreasing the crystallinity of phloretin or phloretin derivative during impregnation in or on mesoporous silica, the dissolution rate in water increased significantly. This higher dissolution rate results in higher bioavailability, and in particular in greater absorption of phloretin or phloretin derivative in the gastrointestinal tract. The more amorphous the phloretin or phloretin derivative, the better the dissolution in water.

In a preferred embodiment of the invention, the method comprises the steps of: iii. providing a mesoporous silica, preferably with a specific surface area, calculated from nitrogen sorption isotherms at liquid nitrogen temperature (77K) according to the Brunauer, Emmett and Teller (BET) theory, of at least 500 m?/g; iv. impregnating said mesoporous silica with phloretin, a phloretin derivative or a combination thereof by solvent impregnation.

The phloretin, phloretin derivative or combination thereof to be loaded onto the silica may be dissolved or suspended in a suitable solvent, and the silica particles may be incubated in the resulting solution for a suitable period of time. Removal of the solvent results in deposition of the phloretin, phloretin derivative or combination thereof on the surface of the silica particles.

In the case of mesoporous silica, the ingredient solution penetrates into the pores of the silica by capillarity and, after removal of the solvent, the ingredient is present in the pores. The preferred solvents are ethanol, methanol, formic acid and isopropyl alcohol. It is clear that the solvent used must be

28 BE2023/5366 sufficiently miscible with phloretin, phloretin derivative or their combination to be impregnated onto silica.

In another aspect, the present invention relates to the composition of the first aspect for solubilizing polyphenols or flavonoids, in particular polyphenols or flavonoids providing aroma or taste. In one embodiment, the present invention relates to a composition comprising a mesoporous silica with a specific surface area, calculated from nitrogen sorption isotherms at liquid nitrogen temperature (77K) by the Brunauer, Emmett and Teller (BET) theory, of at least 500 m2/g, wherein said mesoporous silica is impregnated with polyphenols, flavonoids or mixtures thereof. The inventors have found that the present invention is particularly suitable for solubilizing flavonoids and polyphenols, thereby releasing their flavors, tastes and aromas at a lower concentration and/or at a higher intensity. Advantageously, the present composition can be used in the form of a dry composition, which is advantageous for use in many food formulations.

In a preferred embodiment, the preferred polyphenols are 4-hydroxydihydrochalcones. More preferably, the polyphenols are selected from the following list: 3-(4-hydroxyphenyl)-1-(2-hydroxyphenyl)propan-1-one; 3-(4-hydroxyphenyl)-1-(2,4-dihydroxyphenyl)propan-1-one; 3-(4-hydroxyphenyl)-1-(2,6-dihydroxyphenyl)propan-1-one; 3-(4-hydroxyphenyl)-1-(2,4,6- trihydroxyphenyl)propan-1-one; 3-(4-hydroxy-3-methoxyphenyl)-1-(2-hydroxyphenyl)propan-1-one; 3-(4-hydroxy-3-methoxyphenyl)-1-(2,6-dihydroxyphenyl)propan-1-one; 3-(4-hydroxy-3-methoxyphenyl)-1-(2,4,6-trihydroxyphenyl)propan-1-one; 3-(3,4-dihydroxyphenyl)-1-(2,4,6-trihydroxyphenyl)propan-1-one; salts of the above-mentioned polyphenols; and mixtures of these polyphenols and/or their salts.

In another preferred embodiment, the flavonoids are selected from the following list: quercetin; rutin; kaempferol; hesperidin; naringin; apigenin; luteolin; myricetin; their salts and their mixtures. Most preferably, the flavonoids are selected from the following list: quercetin, rutin and hesperidin, and their salts; and their mixtures. Their mixtures include mixtures of said flavonoids and their salts.

29 BE2023/5366

In a particularly preferred embodiment, the invention relates to a composition comprising a mesoporous silica with a specific surface area, calculated from nitrogen sorption isotherms at liquid nitrogen temperature (77K) by the Brunauer, Emmett and Teller (BET) theory, of at least 500 m2/g, wherein said mesoporous silica is impregnated with a polyphenol or a flavonoid selected from the following list: 4-hydroxydihydrochalcones, preferably phloretin; quercetin; rutin; hesperidin; their salts and mixtures thereof. The inventors have found that these flavoring agents are particularly well suited to combine with mesoporous silica for improved solubilization, in particular because they provide distinct flavor-enhancing effects at low doses, while having very low solubility in water. The inventors have found that the use of these flavoring agents in a composition as described herein allows significantly lower concentrations to be used to obtain the same flavor profile as supplements comprising these flavoring agents available on the market. In another preferred embodiment, said mesoporous silica is impregnated with phloretin and a flavonoid, preferably a flavonoid selected from the following list: quercetin; rutin; hesperidin; their salts and their mixtures.

In another preferred embodiment, the invention relates to a composition comprising a mesoporous silica having a specific surface area, calculated from nitrogen sorption isotherms at liquid nitrogen temperature (77K) according to the Brunauer, Emmett and Teller (BET) theory, of at least 500 m2/g, wherein said mesoporous silica is impregnated with phloretin, glycosylated phloretin or a combination thereof, said glycosylated phloretin being phlorizin and phloretin 3',5'-di-C-glucoside. Preferably, this composition is for use in foods, beverages or nutritional supplements. Another preferred embodiment of the composition is for use in nutrition, oral hygiene, or enjoyment. The composition can be used in dietary supplements, oral hygiene products, or other personal care and wellness products.

A preferred embodiment of the composition comprises phloretin (A) and either no phloridzin or phloridzin in a phloretin/phloridzin ratio of at least 1:3, preferably at least 1:1, more preferably at least 10:1. Phloridzin is known to be bitter and can reduce the sweet taste accentuation of phloretin. Therefore, a high ratio of phloretin to phloridzin is preferred.

30 BE2023/5366

Another preferred embodiment of the invention is a food or beverage product that comprises the composition described above. The food or beverage product may include any type of food or beverage, such as snacks, desserts, juices, teas or coffees. The addition of the composition may improve the taste, odor and nutritional value of the food or beverage, and provide health benefits to the consumer.

Another preferred embodiment of the invention is a food or beverage product comprising the composition described above, wherein the impregnated amount of phloretin, glycosylated phloretin, or their combination is between 0.01 and 150 ppm, preferably between 1 and 100 ppm, more preferably between 5 and 50 ppm, and most preferably between 5 and 25 ppm based on the total weight of the food or beverage. Preferably, the amount of phloretin is between 0.01 and 150 ppm, preferably between 1 and 100 ppm, more preferably between 5 and 50 ppm, and most preferably between 5 and 25 ppm based on the total weight of the food or beverage.

This range gives an indication of the appropriate concentration of the composition in the food or drink product to achieve the desired effects.

The invention is further described by the following non-limiting examples which further illustrate the invention, and are not intended to limit the scope of the invention, nor should they be construed as limiting the same.

EXAMPLES AND/OR DESCRIPTION OF FIGURES

In order to better illustrate the properties of the invention, the following presents, by way of example and without in any way limiting other potential applications, a description of a certain number of preferred applications of the method of examining the condition of the grout used in a mechanical connection based on the invention, in which:

Comparative example 1: crystallinity of a high purity apple extract (>95% phloretin)

Phloretin is extracted from the dried bark of the root of Malus Pumila. Phloretin powder is a crystalline material as shown by the X-ray reflection in Figure 1, which shows the intensity (I) of X-ray reflections from phloretin as a function of 20, where 20 is the angle between the incident ray and the reflected ray.

31 BE2023/5366

Characterization of the amorphous or crystalline state of phloretin was performed by X-ray diffraction (XRD) using a Bruker D2 Phaser diffractometer with Cu-Ka radiation. Samples were analyzed over a range of 6° to 50° (20) with a step of 0.04° at a speed of 2°/min.

Example 2 and comparative example 3

Mesoporous silica (>99%) is provided with the following properties: Mesoporous silica exhibits a high specific surface area, as shown by the nitrogen physisorption curves, where a specific surface area of 680 m2/g was determined from the amount of nitrogen adsorbed. The pore size distribution of mesoporous silica can be observed in Figure 2, which shows the pore volume dV/dW (cm2/g-nm) as a function of pore size (nm).

Mesoporous silica as described above and phloretin as described in Comparative Example 1 are ball milled in a weight ratio of 70/30 to obtain a solid dispersion of phloretin (Example 2) under the following processing conditions: e XQM-2 ball mill from TMAX (China). e Stainless steel balls e Rotation speed: 450 rpm e Container volume: 500 ml e Time: 120 minutes e Single rotation direction (no change of direction during milling)

The solid dispersion based on phloretin obtained by ball milling (Example 2) was characterized by X-ray diffraction and the obtained reflections were compared to a physical mixture of mesoporous silica and pure crystalline material of phloretin. The comparison of the reflection of the two materials can be observed in Figure 3, which shows the intensity (I) of the X-ray reflections of the composition as a function of 20, where 26 is the angle between the incident and the reflected ray.

Characterization of the amorphous or crystalline state of the composition was performed by X-ray diffraction (XRD) using a Bruker D2 Phaser diffractometer with Cu-Ka radiation. The samples were analyzed over a range of 6° to 50° (20) with a step of 0.04° at a speed of 2°/min.

32 BE2023/5366

Comparison of X-ray reflections of the solid dispersion of phloretin (Example 2) and the physical mixture between pure crystalline phloretin material and mesoporous silica (Comparative Example 3) shows that the ball milling process amorphized the phloretin, as the intensities of the X-ray reflections characteristic of phloretin decrease after the ball milling process.

Example 4

Mesoporous silica (>99% SiO2) is provided with the following properties: Mesoporous silica has a high specific surface area, as shown by the nitrogen physisorption curves, with a specific surface area of 517 m2/g, determined from the amount of adsorbed nitrogen. For the solvent impregnation process, the total pore volume is the property of interest in order to determine the maximum amount of active substance that can be incorporated into the pores of mesoporous silica. The pore size distribution of mesoporous silica can be observed in Figure 4, which shows the pore volume dV/dlog(W) (cm3/g-nm) as a function of pore size (nm). Mesoporous silica has a total pore volume of 0.93 cm?/9g.

Mesoporous silica, as described above, is impregnated with phloretin, as described in Comparative Example 1, by solvent impregnation.

A 0.1 g/ml solution of phloretin in ethanol is prepared in a first step. 4.17 ml of this solution are introduced dropwise into 5.0 grams of the mesoporous silica while maintaining manual stirring with a spatula until the material has regained a dry appearance. Ethanol was chosen as the impregnation solvent because of its safety compared to other organic solvents and because the solubility of phloretin in ethanol is relatively high (0.84 mol/kg at 25 °C). — The addition is repeated 3 times (4 additions of 4.17 ml in total), and the product is dried overnight in an oven at 40 °C to completely remove the solvent.

The material obtained after the solvent impregnation process (example 4) was characterized by X-ray diffraction and the obtained reflections were compared to a physical mixture of mesoporous silica and pure crystalline phloretin material (comparative example 3).

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Characterization of the amorphous or crystalline state of the composition was performed by X-ray diffraction (XRD) using a Bruker D2 Phaser diffractometer with Cu-Ka radiation. The samples were analyzed over a range of 6° to 50° (20) with a step of 0.04° at a speed of 2°/min.

Comparison of X-ray reflections of the phloretin-based material obtained by solvent impregnation (Example 4) and the physical mixture between pure phloretin crystalline material and mesoporous silica (Comparative Example 3) also shows an amorphization of phloretin by solvent impregnation, since the intensities of X-ray reflections characteristic of phloretin decrease after impregnation.

Comparison of kinetic release of phloretin

The kinetic release of phloretin from the solid dispersion of phloretin obtained by ball milling (Example 2) was compared with the kinetic release of pure crystalline phloretin material (Comparative Example 1) under the release conditions described below: e Apparatus: European Pharmacopoeia Type II dissolution apparatus (blades) - AT Xtend (Sotax) e Dissolution medium: Ultrapure water e Volume of medium: 900 ml e Rotation speed: 75 rpm e Stirrer: blades e Temperature of medium: 37.0 °C + 0.5 °C e Sampling volume: 5 ml filtered through PTFE membrane (0.45 um) - syringe filter. Fresh medium (ultrapure water) is added to each vial where sampling was performed. e Sampling frequency: 0.25 h; 0.5 h; 1 hour and 2 hours e Phloretin dose per sample: 50 mg

The comparison of the phloretin release kinetics of the two products is shown in Figure 5. The release test shows that the amount of dissolved phloretin for the pure crystalline material (comparative example 1) reaches 55% by weight of dissolved active after 2 hours at 37 °C in water. For the solid dispersion (example 2), this amount reaches about 83% by weight.

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The amount of phloretin released is multiplied by 1.5 when it is in the form of a solid dispersion supported by a mesoporous silica with a high specific surface area (680 m2/g).

Example 6: XRD crystallinity measurement

The crystallinity of the product or the XRD crystallinity of phloretin, phloretin derivative or their combination, as defined herein, was determined using the Segal formula: Crl=(Icy-Iam)/Icry WHERE Icry is the intensity of the peak at a 26 between a of 26° and 27.5° and Ian is the intensity of the baseline at a 29 between 24 and — 25.5°, where 26 is the angle between the incident ray and the reflected ray. The peak intensity was defined as the maximum measured value of the peak measured between the two respective intervals.

The XRD crystallinity of the mixture of Comparative Example 3 was calculated to be 61.8%. The XRD crystallinity of Example 2, the same mixture after intensive ball milling, was calculated to be 6.1%. It is clear that a substantial reduction in the crystallinity of phloretin was achieved.

It is clear that the method according to the invention and its applications are not limited to the examples presented.

Examples 7 to 12: Improvement of the sweet taste of phloretin

The high purity phloretin extract characterized in Comparative Example 1 and the dry dispersion of mesoporous silica and phloretin according to Example 2 were added to a series of sweetened foods and beverages. The respective treats and flavors were evaluated by a test panel (n=10), which compared two food products and was asked to evaluate which, if any, was sweeter.

In Comparative Examples 7 and 8, phloretin was added to a sweetened beverage containing 10% by weight sucrose and 0.3% by weight citric acid.

In Comparative Example 7, 50 ppm of the high purity phloretin extract of Comp. Example 1 was added. In Example 8, 80 ppm of the dry dispersion of silica and phloretin of Example 2 was added. This corresponds to a concentration of 24 ppm phloretin.

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Of the panel members, 5 found no difference between the sweetness of the two drinks; 4 members found Example 8 to be sweeter; 1 member found Comp. Example 7 to be sweeter.

In Comparative Example 9, 24 ppm of the high purity phloretin extract from Comp. Example 1 was added to the same sweetened beverage. The test panel compared Comp. Example 9 and Example 8.

In Comparative Example 10, Example 11 and Comparative Example 12, chewing gums flavored with 1% by weight aspartame and 0.5% by weight menthol were combined with phloretin.

In Comparative Example 10, 100 ppm of high purity phloretin extract from Comp. Example 1 was added. In Example 11, 48 ppm of phloretin was added in the form of the dry dispersion from Example 2. This corresponds to 160 ppm of the dry dispersion, in a ratio of 30/70 between phloretin and silica.

The test panel compared the sweetness of Examples 10 and 11. 9 panelists found no difference in the sweetness of either gum. 1 person found Comp. Example 10 to be sweeter.

In Comparative Example 12, 48 ppm of high purity phloretin extract from Comp. Example 1 was added. The test panel compared the sweetness of — Example 11 and Comp. Example 12. 3 panelists found no difference in the sweetness of either gum. 5 panelists found Example 11 to be sweeter, 2 panelists found Comp. Example 12 to be sweeter.

Claims (18)

36 BE2023/5366 CLAIMS 1. A composition comprising a mesoporous silica having a specific surface area, calculated from nitrogen sorption isotherms at liquid nitrogen temperature (77K) according to the Brunauer, Emmett and Teller (BET) theory, of at least 500 m2/g, wherein said mesoporous silica is impregnated with phloretin, glycosylated phloretin, or a combination thereof, said glycosylated phloretin being phlorizin and phloretin 3',5'-di-C-glucoside. 2. Composition according to claim 1, in which said mesoporous silica has mesopores whose average size, calculated from the nitrogen sorption isotherms at the temperature of liquid nitrogen (77K) according to the Barrett, Joyner and Halenda (BJH) method, is between 1 and 6 nm, preferably between 1 and 3 nm. 3. A composition according to any preceding claim, wherein more than 60% by weight of phloretin or glycosylated phloretin is dissolved in water after 2 hours at 37°C. 4. Composition according to any one of the preceding claims, in which the ratio of the total amount of said phloretin and glycosylated phloretin to said mesoporous silica by weight is between 10/90 and 50/50, preferably between 20/80 and 40/60. 5. Composition form according to any one of the preceding claims, in which said composition comprises said phloretin or said phloretin derivative in an amount of at least 1% by weight relative to the total weight of said composition, preferably at least 10% by weight, more preferably at least 20% by weight. 6. A composition according to any preceding claim, wherein the phloretin or glycosylated phloretin has an XRD crystallinity of less than 50%. 7. A composition according to any preceding claim, wherein the median particle size (d50), as determined by SEM, of the phloretin, phloretin derivative or combination thereof is at most 10 um. 37 BE2023/5366 8. Oral dosage form comprising a composition according to any one of claims 1 to 7. 9. Oral dosage form according to claim 8, comprising said composition in an amount of at least 1% by weight, relative to the total weight of said oral dosage form, preferably at least 10% by weight, more preferably at least 25% by weight, more preferably at least 50% by weight, most preferably at least 80% by weight. 10. Composition according to any one of claims 1 to 7, intended to be used in the treatment and/or prevention of a demyelinating disease. 11. Composition form according to claim 10, in which the demyelinating disease is a neurodegenerative and demyelinating disease of the central nervous system. 12. Composition form according to claim 11, in which the neurodegenerative and demyelinating disease of the central nervous system is multiple sclerosis; in particular progressive multiple sclerosis (PMS), more particularly primary progressive MS (PPMS), secondary progressive MS (SPMS) or relapsing-remitting MS (RRMS). 13.Composition according to claim 10, in which the demyelinating disease is a neurodegenerative and demyelinating disease of the peripheral nervous system. 14. Composition according to claim 13, in which the neurodegenerative and demyelinating disease of the peripheral nervous system is chosen from diabetic neuropathy, Charcot-Marie-Tooth disease or traumatic nerve injury, chronic inflammatory polyradiculoneuropathy, Guillain Barre syndrome. 15.A process for producing a composition, comprising the steps of: ii — providing a mesoporous silica whose specific surface area, calculated from nitrogen sorption isotherms at the nitrogen temperature 38 BE2023/5366 liquid (77K) according to the Brunauer, Emmett and Teller (BET) theory, is at least 500 m2/g; and ii. — impregnating said mesoporous silica with phloretin, a phloretin derivative or a combination thereof. 16. A method according to claim 15, wherein the impregnation is selected from the following list: ball milling, hammer milling, twin screw granulation, certech milling, solvent impregnation, or a combination of these methods, preferably ball milling, solvent impregnation or a combination of these. 17. A method according to claim 15 or 16, wherein during the impregnation in step ii. the particle size d90, as measured by SEM, of the phloretin, the phloretin derivative or their combination, is reduced by at least a factor of 2, preferably by at least a factor of 10. 18. A method according to any one of claims 15 to 17, wherein, during the impregnation in step ii. the XRD crystallinity of the phloretin, the phloretin derivative or their combination is reduced by at least a factor of 1.25, preferably by at least a factor of 1.5, more preferably by at least a factor of 2.