Flavylium cations and electrolyte complexes thereof

Abstract

An electrolyte complex is derived from a reaction mixture comprising an anionic saccharide, or a pharmaceutically acceptable salt thereof; and a flavylium cation, a glycoside thereof, an acylated derivative thereof, a pharmaceutically acceptable salt thereof, or a combination thereof. The flavylium cation may be derived from polyphenol enriched plants including berries. Also disclosed are polyphenol-enriched berry extracts and methods of preparing them. In particular, the disclosed extracts and complexes may be used in the treatment and prevention of metabolic disorders such as obesity, insulin resistance, type (2) diabetes, and polycystic ovarian syndrome.

Description

FLAVYLIUM CATIONS AND ELECTROLYTE COMPLEXES THEREOF CROSS REFERENCE TO RELATED APPLICATIONS This application claims priority to Provisional Application No. 63 / 662,091, filed June 20, 2024, the content of which is incorporated by reference in its entirety. BACKGROUND

[0001] Sucrose octasulfate, a highly sulfated, negatively charged carbohydrate derivative of sucrose, has gained attention in the area of drug delivery because of its unique highly sulfated, polyanionic nature and its ability to form electrolyte complexes and coacervates with cationic materials.

[0002] Orally-administered coacervates derived from sucrose octasulfate are prone to further dissolution and decomposition upon reacting with hydrochloric acid naturally present in the human stomach, which may limit the clinical efficacy and treatment potential of these compounds and potentially increase the undesirable side effects due to the absorption of degradation products into systemic circulation and resulting toxicity.

[0003] Therefore, there is a need for electrolyte complexes that are acid-stable when administered orally, for use as potential pharmacological agents and as delivery agents for other pharmaceutical or herbal preparations. BRIEF DESCRIPTION

[0004] Disclosed herein is an electrolyte complex derived from a reaction mixture comprising an anionic saccharide, or a pharmaceutically acceptable salt thereof; and a flavylium cation, a glycoside thereof, an acylated derivative thereof, a pharmaceutically acceptable salt thereof, or a combination thereof.

[0005] Also disclosed is a pharmaceutical composition comprising a therapeutically effective amount of the above-referenced electrolyte complex, wherein the electrolyte complex is a pharmaceutically active agent.

[0006] Another pharmaceutical composition comprises a therapeutically effective amount of an active ingredient other than the above-referenced electrolyte complex, and excipients comprising the electrolyte complex.

[0007] Also disclosed is a composition comprising a polyphenol or a polyphenol- enriched extract and an additive comprising the above-referenced electrolyte complex.

[0008] Disclosed is a polyphenol-enriched berry extract comprising at least 20 wt% polyphenol, based on the total dry weight of the extract.

[0009] A pharmaceutical composition comprises the above-referenced polyphenol- enriched berry extract and a pharmaceutically acceptable excipient.

[0010] A method for treating or preventing obesity in a subject in need thereof, comprises administering to the patient in need thereof a therapeutically effective amount of the above- referenced electrolyte complex, or the pharmaceutical composition comprising the above- referenced electrolyte complex.

[0011] A method for improving glycemic control in a subject in need thereof, comprising administering to the patient in need thereof a therapeutically effective amount of the above- referenced electrolyte complex, or the pharmaceutical composition comprising the above- referenced electrolyte complex.

[0012] A method for treating or preventing obesity in a patient in need thereof, comprises administering to the patient in need thereof a therapeutically effective amount of the above- referenced polyphenol-enriched berry extract or the pharmaceutical composition comprising the above-referenced polyphenol-enriched berry extract.

[0013] A method for improving glycemic control in a patient in need thereof, comprising administering to the patient in need thereof a therapeutically effective amount of the above- referenced polyphenol-enriched berry extract or the pharmaceutical composition comprising the above-referenced polyphenol-enriched berry extract. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] The following FIGURES are exemplary embodiments wherein the like elements are numbered alike.

[0015] FIG. 1 is a Fourier-transform infrared spectroscopy (FTIR) chromatograph of % transmittance versus wavenumber (inverse centimeters, cm-1) for the purified extract of Example 2.

[0016] FIG.2 is a graph of the FTIR changes of intensity versus wavenumber (cm-1) for the electrolyte complexes of Example 3.

[0017] FIG. 3 is a graph of the glucose tolerance test of blood glucose (milligrams per deciliter, mg / dl) versus time (minutes, min) for the materials as described in Example 4.

[0018] FIG.4 is a graph of the weight loss in obese mice of weight loss (%) versus days of administration (days) as described in Example 5.

[0019] FIG. 5 is a photograph of the cross-linked electrolyte complex product of Example 7.

[0020] FIG. 6 is a graph of weight loss in obese mice of weight loss (%) after two weeks of treatment as described in Example 9. DETAILED DESCRIPTION

[0021] The details of one or more embodiments of the presently-disclosed subject matter are set forth in this document. Modifications to embodiments described in this document, and other embodiments, will be evident to those of ordinary skill in the art after a study of the information provided in this document. The information provided in this document, and particularly the specific details of the described exemplary embodiments, is provided primarily for clarity of understanding and no unnecessary limitations are to be understood therefrom. In case of conflict, the specification of this document, including definitions, will control.

[0022] Chemicals based on the sucrose octasulfate have been known and used to treat gastrointestinal disorders since 1968, with sucrose octasulfate-aluminum hydroxide complex (sucralfate) being the particular example of such complex. In the recent years, fresh attention has been re-drawn to the complex coacervates formed upon reacting the sucralfate with hydrochloric acid, and potential use of such coacervates as bioactive chemicals which could be used as orally deliverable pharmaceuticals. Coacervation is a process where liquid–liquid phase separation occurs between two oppositely charged polymers (or polyelectrolytes), forming a dense, polymer-rich phase (the coacervate) and a dilute supernatant. This allows encapsulation of active compounds or formation of bioadhesive gels. Sucrose octasulfate, a highly sulfated, negatively charged carbohydrate derivative of sucrose, has gained attention in the area of drug delivery because of its unique highly sulfated, polyanionic nature and its ability to form electrolyte complexes and coacervates with cationic materials (e.g., polycations, multivalent metal ions, and cationic proteins and peptides). These properties allow it to be used as a bioadhesive, protective matrix, or carrier system for various drugs. Sucrose octasulfate is best known as the active polyanionic component in the drug sucralfate, and for its mucosal protective, wound healing, and bioadhesive properties. For example, sucralfate is a gastroprotective medication used primarily to treat and manage peptic ulcers and other mucosal injuries of the gastrointestinal (GI) tract. Aluminum hydroxide makes the complex poorly soluble, so sucralfate remains localized in the stomach, without systemic absorption. At pH < 4 (like in the stomach), sucralfate forms a viscous, sticky gel.

[0023] However, sucrose octasulfate alone generally does not form a stable hydrogel without a complementary crosslinking agent like aluminum hydroxide or another multivalent cation / polycation. crosslinkers, bridging multiple SOS molecules and forming a stable gel. Alternatively, SOS can form coacervates or gels when combined with cationic polymers (like chitosan or poly-L- lysine), which provide multiple positive charges to crosslink SOS.

[0024] Oral delivery implies that the coacervates have to pass through the stomach, which naturally produces hydrochloric acid. The chemical nature of said coacervates makes them prone to further dissolution and decomposition upon reacting with hydrochloric acid naturally present in the human stomach, which may limit the clinical efficacy and treatment potential of these compounds and potentially increase the undesirable side effects due to the absorption of degradation products into systemic circulation and resulting toxicity.

[0025] The present inventors have found that the disclosed compositions can modulate the solubility of sucrose sulfate-aluminum hydroxide coacervate complexes in hydrochloric acid while allowing to maintain sufficient hydration and formation of hydrogels. These compositions include electrolyte complexes, which have improved stability in the acidic environment (e.g., GI tract following oral administration), which may increase the efficacy against certain disorders and reduces the risk of exposure of the organs and tissues of the body to the degradation products of the parent compounds otherwise absorbed into systemic circulation.

[0026] The disclosed electrolyte complexes may form electrolyte networks, wherein the electrolyte networks may be crosslinked. This results in a combination of properties typical of both hydrogels and ionic materials. The electrolyte complexes may be chemically crosslinked (e.g., covalent bonds) or physically crosslinked (e.g., ionic interactions, hydrogen bonds).

[0027] The disclosed electrolyte complexes may be derived from a reaction mixture including an anionic saccharide, or a pharmaceutically acceptable salt thereof; and a flavylium cation, a glycoside thereof, an acylated derivative thereof, a pharmaceutically acceptable salt thereof, or a combination thereof. Advantageously, the electrolyte complex possesses substantially different chemical and / or physical properties from the anionic saccharide and the flavylium cation, and can be separated from the residual reactants. The electrolyte complex may be crosslinked. Crosslinking may be via formation of a chemical bond, including, but not limited to covalent bonds, coordinated bonds, hydrogen bonds, Van der Waals interactions, and ionic or electrostatic bonds. Not wishing to be bound by theory,since the anionic saccharide and the flavylium cation carry opposite charges, it is likely that the crosslinking is electrostatic or ionic.

[0028] The electrolyte complexes may be prepared by combining the anionic saccharide and the flavylium cation in aqueous solution with stirring. The weight ratio of the anionic saccharide to the flavylium cation in the reaction mixture may be adjusted to modulate the solubility of the electrolyte complex. For example, the weight ratio may range from about 1:1 to about 1000:1. Within this range, the ratio may range from about 1:1 to about 500:1, about 2:1 to about 500:1, about 3:1 to about 500:1, about 4:1 to about 500:1, about 5:1 to about 500:1, about 10:1 to about 500:1, about 50:1 to about 500:1, about 100:1 to about 500:1, or about 100:1 to about 1000:1.

[0029] The anionic saccharides of the disclosed complexes may encompass anionic monosaccharides, anionic disaccharides, anionic polysaccharides, and combinations thereof. As used herein, “anionic saccharide” encompasses the free anion and pharmaceutically acceptable salts thereof. The anionic saccharides include at least one anionic group, e.g.,carboxylate (– , phosphate (– , and sulfate (– . The anionic saccharidemay include at least two, at least three, at least four, at least five, at least six, at least seven, or at least eight anionic groups. The anionic groups may include carboxylate groups, phosphate groups, sulfate groups, or a combination thereof. In some aspects, the anionic groups include carboxylate groups and / or sulfate groups. In some aspects, the anionic groups include carboxylate groups. In some aspects, the anionic groups include sulfate groups. Non-limiting examples of anionic monosaccharides include glucoronic acid, galacturonic acid, iduronic acid, mannuronic acid, sulfated glucose, sialic acid, rhamnose sulfate, and 2-keto-3-deoxy- octulosonic acid. Non-limiting examples of anionic disaccharides include sucrose sulfate (1-8 sulfate groups), cellobiose sulfate (up to 8 sulfate groups), chondroitin sulfate and polysulfate derivatives thereof, and maltose sulfate and polysulfate derivatives thereof (e.g., maltose heptasulfate).

[0030] The anionic saccharides may include an anionic monosaccharide, or a pharmaceutically acceptable salt thereof, and optionally, an anionic disaccharide, an anionic polysaccharides, a combination thereof, or a pharmaceutically acceptable salt thereof. The anionic saccharides may include an anionic monosaccharide only. The anionic saccharides may include an anionic disaccharide only. The anionic saccharides may include an anionic polysaccharide only. The anionic saccharides may include an anionic monosaccharide and an anionic disaccharide. The anionic saccharides may include an anionic monosaccharide and an anionic polysaccharide. The anionic saccharides may include an anionic disaccharide and ananionic polysaccharide. The anionic saccharides may include an anionic disaccharide and anionic polysaccharide. When two anionic saccharides are present, the weight ratio may be, for example, 1:99 to 99:1. Within this range, the weight ratio may be 1:95 to 95:1, 1:90 to 90:1, 1:5 to 5:1, 1:4 to 4:1, 1:3 to 3:1, 1:2 to 2:1, or 1:1.

[0031] Anionic polysaccharides may include an anionic mucopolysaccharide / an anionic glycosaminoglycan, an anionic mucilage, a combination thereof, or a pharmaceutically acceptable salt thereof. Glycosaminoglycans (GAGs) also referred to as mucopolysaccharides are long unbranched polysaccharides containing a repeating disaccharide unit, where each unit includes at least one sugar bearing a negative charge,typically due to the presence of carboxylate (– or sulfate (– groups. Non-limiting examples of anionic mucopolysaccharides may include heparin, heparin sulfate, chondroitin sulfate, dermatan sulfate, hyaluronic acid, and keratin sulfate. The disaccharide units may include either of two modified sugars, N-acetylgalactosamine (GalNAc) or N- acetylglucosamine (GlcNAc), and a uronic acid such as glucuronate or iduronate. The hyaluronates may be composed of D-glucuronate and GlcNAc. The dermatan sulfates may be composed of D-glucuronic acid (GlcA) or L-iduronate (IdoA) and GalNAc-sulfate. Heterogeneity in dermatan sulfate results from varying degrees of O-sulfation and from the presence of the two uronic acids. Chondroitin sulfates may be composed of D-glucuronate and GalNAc-6 (or 4)-sulfate. Heparin and heparin sulfates may be composed of D- glucuronate-2-sulfate and N-sulfo-D-glucosamine-6-sulfate (heparins have less sulfate than heparins). Keratin sulfates may be composed of galactose and galactose-6-sulfate and GlcNAc-6-sulfate. Chondroitin sulfate is a sulfated glycosaminoglycan (GAG). A chondroitin chain may have over 100 individual sugars, each of which can be sulfated in variable positions and quantities.

[0032] An anionic mucilage is a type of plant-derived polysaccharide that is gel-like, highly water-absorbing, and carries a net negative charge due to the presence of acidicfunctional groups—typically uronic acids (carboxyl groups, – sulfate groups (–Generally, mucilage can be obtained from several plants or their different parts such as, for example, aloe vera, salvia hispanica seeds, cordia dichotoma, basella alba, plantago psyllium, cyamopsis tetragonoloba, Cactaceae, abelmoschus esculentus, trigonella foenum- graecum, moringa oleifera, laminaria, ascophyllum, and linum usitatissimum. Plant-derived mucilage may be extracted from the plant using maceration, followed by precipitation. Structurally, mucilage (a complex of polymeric polysaccharide) is mainly composed of carbohydrates with highly branched structures that are derived from monomer units includingL-arabinose, D-xylose, D-galactose, L-rhamnose, and galacturonic acid. Anionic mucilage can also be obtained from fruits, particularly those that contain pectic polysaccharides oracidic heteropolysaccharides rich in uronic acids (which provide the anionic –

[0033] Anionic polysaccharides (similar to anionic mono and di-saccharides) may be derived from natural sources or may be synthetic (or semi-synthetic). Non-limiting examples of anionic polysaccharides include carrageenans, alginates, agar, pectins, modified pectins, gellan gum, xanthan gum, furcelaran, cellulose derivatives, particularly carboxymethyl cellulose (CMC) and cellulose sulfate, dextran sulfate, modified starches, exo polysaccharides, polyacrylic acid, and combinations thereof. In an aspect, the anionic anionic polysaccharide includes xanthan gum, alginate (e.g., sodium alginate), or a combination thereof. Xanthan gum is a natural, anionic polysaccharide widely used as a thickener, stabilizer, and suspending agent in food, pharmaceutical, cosmetic, and industrial products. It is produced by the fermentation of sugars by the bacterium Xanthomonas campestris. Xanthan gum includes glucoronic acid and pyruvic acid residues, which impart the anionic character and water solubility. Alginate (or alginic acid) is a natural, anionic polysaccharide primarily extracted from brown seaweeds such as Laminaria, Macrocystis, and Ascophyllum. Alginate includes guluronic acid and mannuronic acid residues, which impart the anionic character and water solubility.

[0034] The electrolyte complex may be derived from an anionic saccharide including a sucrose sulfate (degree of sulfation 1-8) and a flavylium cation. The resulting electrolyte complex may have the following Formula I: (C12H6(-OSO3)m1)•(R1)n)m2• (C15H11OR2)+. The designation “m1” may be from 1 to 8 and “m2” may be from -1 to -8. R1can be a proton, a monovalent metal cation, a positively charged metal hydroxide such as (Ca(OH)+Mg(OH)+, Fe(OH)n+, and Al(OH)n+, with elemental metal content of about 0.1 to about 15%, about 0.5 to about 15%, about 1 to about 15%, about 5 to about 15%, about 1 to about 10%, about 5 to about 10 (as measured by inductively coupled plasma mass spectrometry, ICPMS) and n for Me(OH)+in the range of 0 < n < 15. The Me+may form chemical bonds with the sulfate and also form secondary coordinated shells. For example, a description of said sucrose octasulfate-aluminum salt is given by US11,524,024 B2, US10,716,802 B2, US11,666,597B2, each incorporated by reference in their entirety herein. A higher Al(OH)2+content may be disadvantageous due to the tendency to form water-insoluble and non- hydratable salts with sucrose sulfate, which is observed for 15-25 weight % elemental Al preparations. R2 can be a proton, a hydroxyl, a glycoside, an acyl group, an alkyl group (e.g., methyl), or a salt-forming cation.

[0035] The flavylium cation may be the core structure (2-phenylchromenylium (or 2- phenyl-1-benzopyrylium)) of anthocyanins. The flavylium cation exists under acidic pH and is involved in a dynamic system of interconverting species in response to changes in pH, including quinonoidal bases (pH 4-5), hemiketals, and chalcones (pH 7).

[0036] Flavylium cations may have a basic structure as shown in Formula II.Formula II

[0037] Typical examples are: cyanidin (hydroxylated at positions delphinidin (hydroxylated at positions at positions ) peonidin

[0038] Flavylium cations may include water-soluble glycosides of polyhydroxyl and polymethoxyl derivatives of 2-phenylbenzopyrylium or flavylium salts. Individual flavylium cations may differ in the number of hydroxyl groups present in the molecule, the degree of methylation of these hydroxyl groups, the nature, number and location of sugars attached to the molecule and the number and the nature of aliphatic or aromatic acids attached to the sugars in the molecule. Hundreds of flavylium cations have been isolated and chemically characterized by spectrometric tools. Cyanidins and their derivatives are the most common flavylium cations present in vegetables, fruits and flowers.

[0039] Flavylium cations share a basic carbon skeleton in which hydrogen, hydroxyl or methoxyl groups can be found in six different positions as noted above. In fruits and vegetables, six basic flavylium cations predominate, differing both in the number of hydroxyl groups present on the carbon ring and in the degree of methylation of these hydroxyl groups. The identity, number and position of the sugars attached to the carbon skeleton are also variable; the most common sugars that can be linked to carbon-3, carbon-5 and, sometimes, carbon-7, are glucose, arabinose, rhamnose or galactose. On this basis, it is possible to distinguish monosides, biosides and triosides.

[0040] One or more of the hydroxyl groups of the flavylium cations may be acylated. Typical acylating agents are caffeic, ferulic, sinapic and p-coumaric acids, although aliphaticacids such as acetic, malic, malonic, oxalic and succinic acids can also occur. Combinations of acids are possible.

[0041] Flavylium cations, due in part to the nature of their chemical structure, tend to be unstable and susceptible to degradation. Additionally, the stability of flavylium cations is affected by pH, storage over a period of months, storage temperature, presence of enzymes, light, oxygen, and the presence of proteins, flavonoids and minerals. More particularly, the bioavailability of flavylium cations is low due to their sensitivity to changes in pH. Flavylium cations are generally stable at pH values of 3.5 and below, and are therefore stable under stomach conditions. However, they degrade at higher pH values, such as those more typical for the intestinal tract (pH of 7) and thus beneficial absorption and nutritional value is greatly reduced.

[0042] The flavylium cations may be represented by Formula III. Non-limiting examples of common flavylium cations are included in Table 1 below.

[0043] Flavylium cations may be classified by the number of glycosyl units they contain. Monoglycosides include one saccharidic moiety, which is primarily attached to the 3- hydroxyl group of the aglycon. Diglycosides generally contain two monosaccharides at the 3 and 5 hydroxy positions and occasionally at the 3 and 7 hydroxyl positions. Triglycosideshave attachment generally where there are two units at the 3 position and one at the C-5 or C- ble. In particular, common glycosides of the flavylium cations include glucose, rhamnose, galactose, arabinose and xylose. The di- and trisaccharides found most often in flavylium cations are rutinose, sophorose, sambubiose and glucorutinose. Non-limiting examples of glycosides that can be attached to the flavylium cation can include glucose, galactose, rhamnose, arabinose, xylose, glucuronic acid, sophorose, sambubiose, rutinoside (rhamnose + glucose), neohesperidose (rhamnose + glucose in a different linkage), and gentiobiose.

[0044] The flavylium cation may be derived from anthocyanin-rich, water-soluble purified berry extracts. Anthocyanin-rich berries may be characterized by their red, blue, purple, and black hues. Anthocyanins are a subclass of flavonoids and are known for their antioxidant, anti-inflammatory, and vasoprotective properties. Flavylium cations may be derived from extracts derived from plant sources. Extracts encompass flavylium cation- containing materials obtained from plant sources, such as leaves, twigs, bark, roots, stern, seeds, flowers, berries, and fruit. There are various conventional methods for the extraction of flavylium cations known to those of skill in the art. Some of these methods are described in, for example, U.S. Pat. No. 5,817,354; U.S. Pat. No. 5,200,186; U.S. Pat. No. 5,912,363; U.S. Pat. No. 4,211,577; U.S. Pat. No. 4,302,200 (each incorporated herein by reference). Typically, the extract is concentrated by various methods to provide a solution enriched in flavylium cations. For example, ultrafiltration can be used to remove unwanted components. The retained liquid from the filtration can be stored as a liquid or, for example, can then be further concentrated into a powder by spray drying, freeze drying, flash drying, fluidized bed drying, ring drying, tray drying, vacuum drying, radio frequency drying or microwave drying. Commercially available flavylium cations (or cation precursors) may also be used.

[0045] Conventionally prepared extracts can be further purified by one or more methods known in the art, such as chromatography, gel chromatography, high performance liquid chromatography, crystallization, affinity chromatography, partition chromatography and the like. Identification of the particular flavylium cations can be accomplished by methods know to those skilled in the art and includeNMR, chemical degradation, chromatography and spectroscopy, especially homo- and heteronuclear two-dimensional NMR techniques for the characterization of the isolated flavylium cations.

[0046] Non-limiting examples include fruits, vegetables, flowers and other plants that are good sources of compounds comprising a flavylium cation include: Acer macrophyllum, Acer platanoides, acerola, Ajuga reptans, apple, apricot, Artict bramble, avocado, banana,barberry, barley, Begonia semperfiorens, Bellis perennis, Bletilla striata, bilberry, black beans, black soybeans, black, blue and purple potatoes, blackberry, blueberry, bog whortleberry, boysenberry, buckwheat, cacao, Camellia sinensis, canarygrass, Caucasian blueberry, Chimonanthus praecox, celery, Cerasus avium, cherry, cherry laurel, chicory, chive, chokeberry, Cornelian cherry, cornflower, cotoneaster, cowberry, cranberry, crowbeny, chrysanthemum, Cynomorium coccineum, Dahlia variabilis, danewort, deerberry, Dendrobium, dwarf dogwood, Echinacea purpea, eggplant, elderberry, fababean, Fatsia japonica, feijoa, fig, garlic, gerbera, ginseng, Globe artichoke, gooseberry, grapes, guava, hawthorn, hibiscus or roselle, Hibiscus Sabdaiffa, highbush blueberry, hollyhock, honeysuckle, Ipomoea purpurea, Iris ensata, Java plum, Jerusalem artichoke, kokum, Laeliocattleya, lentil, loganberry, lupine, lychee, maize, mango, mangosteen, maqui, Matthiola incana, meconopsis, Metrosideros excelsa, millet, mountain ash berry, mulberry, myrtle berry, olive, onion, orange, ornamental cherry, passion fruit, pea, peach, peanut, pear, perilla, petunia, Phalaenopsis, Phalsa, Pharbitis, Pineapple, pistachio, plum, pomegranate, Phragmites australis, purple carrot, quince, rabbiteye blueberry, radish, red and black currant, red and black raspberry, red cabbage, rice, rhubarb, rosehip, rye, saffron, sarracenia, sheepberry, Sophronitis coccinea, sorghum, sparkleberry, strawberry, Fragada Vesca, sugarcane, sunflower, sweet cherry, sweet potato, tamarillo, tamarind, taro, tart cherry, Tulip greigii, turnip, water lily, Weigela, wheat, wild rice, Verbena hybrida, yam and mixtures thereof.

[0047] Although there are literally thousands of extracts containing flavylium cations, particularly suitable examples of extracts containing flavylium cations include berries. Berries that are sources of flavylium cations generally include blueberries, bilberries, blackberries, elderberries, black raspberries, cherries, grapes, pomegranate, black currant, purple fig, red raspberries, chokeberries, cranberries, strawberries, acai berries, sloe berries, and mulberries. Berries with the highest amounts of flavylium cations are typically the darker-colored berries, e.g., blueberries, bilberries, blackberries, cherries, grapes, pomegranate, black currant, purple fig, elderberries, black raspberries, chokeberries, cranberries, acai berries, sloe berries, and mulberries.

[0048] The presently-disclosed subject matter includes a berry extract enriched in polyphenol content. Berries are a rich source of phytochemicals, especially phenolic compounds. Polyphenols derived from berries include flavonoids (anthocyanins, flavonols, and flavan-3-ols), condensed and hydrolyzable tannins, phenolic acids, stilbenes, and lignans. Concentrations of the respective polyphenols vary according to species, genotype,environmental conditions, degree of ripeness, cultivar, cultivation site, processing, and storage of the fruit. The polyphenols may have biological activity, such as, e.g., antioxidant, anti-inflammatory, antimicrobial, anticancer, and antidiabetic activities. Therefore, in addition to forming an electrolyte complex, the flavylium cation may exert therapeutic effects.

[0049] In addition to these beneficial effects of polyphenols including anthocyanins, enrichment of the polyphenols of the extracts are preferable for obtaining sufficient concentration of the compounds capable of forming a flavylium cation (e.g., anthocyanins) in the reaction mixtures for preparing the electrolyte complexes, reducing the level of impurities and contaminants in the product, avoiding undesirable reaction products, providing better control of the reaction and reducing the quantity of the extract used (cost savings). This is in contrast to conventional berry extracts, which include about 3 wt% polyphenol content.

[0050] The present inventors found that by macerating the berries in water solvent in the presence of enzymes, that the berry extract could be enriched in polyphenol content. As such, the polyphenol-enriched berry extract may be substantially free of organic solvents, such as alcohol solvents. Maceration of fruits is a process where fruit is softened and broken down to release its juices, flavors, sugars, and pigments. Maceration may be accomplished by soaking in liquids. Conventional approaches for preparing berry extracts from whole berries utilize alcohol (e.g., 40% ethanol in water). Other conventional approaches use berry powder, which is combined with an alcohol solvent (e.g., ethanol in water (70%) or methanol in water (80%)) and heated gently. In the present methods, the berries are soaked in water in the presence of pectolytic enzymes at e.g., room temperature, to provide a macerated fruit mixture. The mixture is filtered, and the filtrate may be purified by chromatography (e.g., SepaBeads). The extract may be eluted with aqueous alcohol and concentrated to remove solvent. By using water instead of aqueous alcohol, the filtrate obtained after filtering the macerated fruit mixture may be enriched in water-soluble polyphenols. This is in contrast to conventional methods that use aqueous alcohol, where polyphenols that are soluble in alcohol, but have limited or poor solubility in water are present to a greater extent in the liquid phase of the macerated fruit mixture. For example, aglycone forms of flavonols, proanthocyanidines, tannins, and stilbenes have poor solubility in water, whereas anthocyanins are highly soluble in water. In particular, by using water instead of water / alcohol mixtures during maceration, the liquid phase of the macerated fruit mixture may be enriched in anthocyanins. This is an advantage in preparing the disclosed electrolyte complexes. Not wishing to be bound by theory, but the anthocyanins form flavylium cationsand the other polyphenols tend to not form stable cations, if at all. Therefore, berry extracts enriched in anthocyanins provide a greater amount of flavylium cation, thus increasing the yield of the electrolyte complexes. For example, using the disclosed method, the liquid phase of macerated sloe berries would be expected to be enriched in anthocyanins due to the high water solubility of anthocyanins and have reduced amounts of tannins, stilbenes, and hydroxycinnamic acid, given their poor water solubility. Although water is preferred as a solvent, the disclosed extraction (e.g., maceration) methods may be carried out in the presence of a co-solvent, such as, for example, an alcohol (e.g., methanol, ethanol, or a combination thereof) such that the solvent mixture comprises at least 50% water, at least 60% water, at least 70% water, at least 80% water, at least 90% water, at least 95% water, or at least 99% water. In some aspects, the solvent is water.

[0051] Measuring polyphenol content—especially in berries, juices, or plant extracts— can be done using both simple spectrophotometric and advanced chromatographic methods. The most common lab-scale method is the Folin–Ciocalteu (F–C ) assay, which gives a good total phenolic content (TPC) estimate. The F–C assay is a colorimetric assay. The oxidizing F–C reagent reacts with reducing agents (antioxidants) to form a soluble, vividly blue complex. The precise chemical structure of the F–C reagent is unknown; however, it is described as a complex mixture of phosphotungstic and phosphomolybdic acids that is reduced throughout the assay to produce a blue chromophore with a maximum absorbance at 765 nm. The reduction of the anionic derivatives of phosphotungstic and phosphomolybdic acids by antioxidants causes a color shift from yellow to blue, and the magnitude of the color shift when the reaction is complete is directly proportional to the reducing activity of the phenolic compounds. The reducing capacity of an antioxidant is frequently measured as gallic acid equivalents (GAE).In more detail, the transfer of electrons from phenolic compounds to phosphomolybdic and phosphotungstic acid complexes in an alkaline solution creates blue complexes that are detected spectroscopically at about 760 nm. To determine the wt% total phenolic content, the total phenolic content (TPC) measured via the F–C assay, expressed as mg gallic acid equivalents (GAE) per gram of dried extract (i.e., dry mass) and multiply by 100%. Although suitable for determining the total polyphenol content of a sample, the F–C assay does not distinguish among individual polyphenols. To distinguish among the individual polyphenols, high-performance liquid chromatography (HPLC) may be used and peaks may be compared to reference samples of the polyphenols to determine the polyphenols present in the extract.

[0052] The presently-disclosed subject matter includes a polyphenol-enriched berry extract comprising at least 20 wt% polyphenol content, based on the total dry weight of the extract. Within this range, the polyphenol-enriched berry extract includes at least 25 wt%, at least 30 wt%, at least 35 wt%, at least 40 wt%, at least 45 wt%, at least 50 wt%, at least 55 wt%, at least 60 wt%, at least 65 wt%, at least 70 wt%, at least 75 wt%, at least 80 wt%, at least 85 wt%, at least 90 wt%, or at least 95 wt%. The upper limit for phenolic content may include up to 100 wt%, less than or equal to 100 wt%, less than or equal to 95 wt%, less than or equal to 90 wt%, or less than or equal to 85 wt%. Any of the foregoing upper and lower limits can be combined to make a range. For example, the polyphenol-enriched berry extract includes 20-100 wt%, 30-100 wt%, 35-95 wt%, 35-90 wt%, or 40-90 wt% polyphenol content.

[0053] In addition to polyphenols, the polyphenol-enriched berry extract may include 0- 20 wt% carbohydrate content. Within this range, the polyphenol-enriched berry extract includes 0-15 wt%, 0-10 wt%, 0-5 wt%, or 0-1 wt% carbohydrate content.

[0054] Berries differ in their anthocyanin content. Black currants have greater than 50% to 97% anthocyanins as compared with total polyphenol content, whereas blackberries have about 30 to 50% anthocyanins as compared with total polyphenol content. The polyphenol- enriched berry extract of the present disclosure may be enriched in anthocyanins as compared to the anthocyanin content of the berry. Blueberries, for example, may include anthocyanin as 20-60% of the total polyphenol content. Despite black currants having a higher anthocyanin content than blueberries, they are not widely available and are typically much more expensive. Therefore, it would be an advantage if the anothocyanin content could be enriched in berry extracts derived from widely available, less expensive berries.

[0055] Using the disclosed methods, the anthocyanin content may be enriched to greater than the anthocyanin content of the unprocessed berry. For example, if the blueberry contains 60% anthocyanin as compared with the total polyphenol content, the anthocyanin content of the blueberry extract may be increased to at least 65%, at least 70%, at least 75%, or at least 80% of the total polyphenol content, as determined by HPLC. In some aspects, using the disclosed methods, the anthocyanin content of the extracts (e.g., anthocyanin-enriched extracts) may be enriched to a value greater than the anthocyanin content of the unprocessed berry, e.g., at least 5% greater, at least 10% greater, at least 15% greater, at least 20% greater, at least 25% greater, at least 30% greater, at least 35% greater, at least 40% greater, at least 45% greater, at least 50% greater, at least 55% greater, at least 60% greater, at least 65% greater, or at least 70% greater than the anthocyanin content of the unprocessed berry, basedon the total polyphenol content. In some aspects, using the disclosed methods, the wt% of anthocyanin based on the total polyphenol content of the extracts may be at least 50%, at least 55%, at least 60%, at least 65%, at least 70%, at least 80%, at least 85%, at least 90%, or at least 95%.

[0056] The polyphenol-enriched (e.g., anthocyanin-enriched) berry extracts may be incorporated into an herbal composition for various culinary applications (e.g., flavoring agent for food and beverages). In a particular aspect, a berry infused alcohol (e.g., vodka, gin) comprises the polyphenol-enriched berry extract.

[0057] The disclosed subject matter includes pharmaceutical composition comprising an electrolyte complex. In some pharmaceutical compositions, the disclosed electrolyte complexes may act as an excipient (e.g., carrier) for a pharmaceutically active agent. The pharmaceutically active agent may be any pharmaceutically active agent known in the art suitable for oral administration. In some aspects, the pharmaceutical compositions include a therapeutically effective amount of a pharmaceutically active agent and excipients comprising the electrolyte complex. The pharmaceutical compositions may include excipients other than a electrolyte complex. A pharmaceutically acceptable excipient includes both one and more than one such excipient. Pharmaceutically acceptable excipients include, but are not limited to, one or more of a binder, a release control agent, a disintegrant, a diluent, or a lubricant. The excipients may further include a glidant, a solvent, a viscosity agent, an emulsifier, a buffer, a bulking agent, a coloring agent, a taste-improving agent, a flow agent, an absorbent, but are not limited thereto.

[0058] The disclosed electrolyte complexes may be used as an additive (e.g., stabilizing agent) in a composition that includes a polyphenol or a polyphenol-enriched extract.

[0059] Berry-derived polyphenols have shown remarkable effects on different types of cancer, including colorectal, breast, esophageal, and prostate cancer. Moreover, certain metabolic disorders such as diabetes and atherosclerosis were also managed by berry-derived polyphenols through different mechanisms. Studies have shown that polyphenols from berries are a promising source of bioactive compounds capable of modulating the intestinal microbiota, and therefore managing cancer and associated metabolic diseases. Therefore, the disclosed polyphenol-enriched berry extracts and electrolyte complexes incorporating the flavylium cation derived from these polyphenol-enriched berry extracts may be used in methods of treating or preventing disease. Non-limiting applications include treatment of insulin resistance, gastrointestinal bloating, flatulence, dyslipidemia, polycystic ovarian syndrome (via glycemic control), fatty liver disease, irritable bowel syndrome, and smallintestinal bacterial overgrowth. The disclosed compositions may also be used to enhance exercise / athletic performance.

[0060] The electrolyte complex may be administered as a pharmaceutically active agent to treat or prevent obesity, and / or to improve glycemic control. A method for treating or preventing obesity in a patient in need thereof comprises administering to the patient in need thereof a therapeutically effective amount of a electrolyte complex, or administering to the patient in need thereof a pharmaceutical composition including a therapeutically effective amount of a electrolyte complex. A method for improving glycemic control in a patient in need thereof, comprises administering to the patient in need thereof a therapeutically effective amount of a electrolyte complex, or administering to the patient in need thereof a pharmaceutical composition including a therapeutically effective amount of a electrolyte complex.

[0061] While the terms used herein are believed to be well understood by those of ordinary skill in the art, certain definitions are set forth to facilitate explanation of the presently-disclosed subject matter.

[0062] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of skill in the art to which the presently disclosed subject matter(s) belong.

[0063] All patents, patent applications, published applications and publications, GenBank sequences, databases, websites and other published materials referred to throughout the entire disclosure herein, unless noted otherwise, are incorporated by reference in their entirety.

[0064] As used herein, the abbreviations for any protective groups, amino acids and other compounds, are, unless indicated otherwise, in accord with their common usage, recognized abbreviations, or the IUPAC-IUB Commission on Biochemical Nomenclature (see, Biochem. (1972) 11(9):1726-1732).

[0065] Although any methods, devices, and materials similar or equivalent to those described herein can be used in the practice or testing of the presently-disclosed subject matter, representative methods, devices, and materials are described herein.

[0066] The present application can “comprise” (open ended) or “consist essentially of” the components of the presently disclosed subject matter as well as other ingredients or elements described herein. As used herein, “comprising” is open ended and means the elements recited, or their equivalent in structure or function, plus any other element or elements which are not recited. The terms “having” and “including” are also to be construed as open ended unless the context suggests otherwise.

[0067] The terms "a" and "an" do not denote a limitation of quantity, but rather denote the presence of at least one of the referenced item.

[0068] Unless otherwise indicated, all numbers expressing quantities of ingredients, properties such as reaction conditions, and so forth used in the specification and claims are to be understood as being modified in all instances by the term “about”. Accordingly, unless indicated to the contrary, the numerical parameters set forth in this specification and claims are approximations that can vary depending upon the desired properties sought to be obtained by the presently-disclosed subject matter.

[0069] As used herein, the term “about,” when referring to a value or to an amount of mass, weight, time, volume, concentration or percentage is meant to encompass variations of in some embodiments ±20%, in some embodiments ±10%, in some embodiments ±5%, in some embodiments ±1%, in some embodiments ±0.5%, in some embodiments ±0.1%, in some embodiments ±0.01%, and in some embodiments ±0.001% from the specified amount, as such variations are appropriate to perform the disclosed method.

[0070] Recitation of ranges of values are merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein. The endpoints of all ranges are included within the range and independently combinable. All methods described herein can be performed in a suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or language indicating an example (e.g. , "such as"), is intended merely for illustration and does not pose a limitation on the scope of the presently disclosed subject matter unless otherwise claimed. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the presently disclosed subject matter.

[0071] "Pharmaceutically acceptable salts" includes compounds modified by making acid or base addition salts thereof, and further refers to pharmaceutically acceptable solvates, including hydrates, and co-crystals of such compounds and such salts. Examples of pharmaceutically acceptable salts include, but are not limited to, mineral or organic acid addition salts of basic residues such as amines; alkali or organic addition salts of acidic residues; and the like, and combinations comprising one or more of the foregoing salts. For example, non-toxic acid salts include those derived from inorganic acids such as hydrochloric, hydrobromic, sulfuric, sulfamic, phosphoric, nitric and the like; other acceptable inorganic salts include metal salts such as sodium salt, potassium salt, cesium salt,and the like; and alkaline earth metal salts, such as calcium salt, magnesium salt, and the like, and combinations comprising one or more of the foregoing salts. Pharmaceutically acceptable organic salts includes salts prepared from organic acids such as acetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, pamoic, maleic, hydroxy maleic, phenylacetic, glutamic, benzoic, salicylic, mesylic, esylic, besylic, sulfanilic, 2- acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, isethionic, HOOC-(CH2)n-COOH where n is 0-4, and the like; A “pharmaceutically acceptable excipient” means an excipient that is useful in preparing a pharmaceutical composition that is generally safe, non-toxic and neither biologically nor otherwise undesirable, and includes an excipient that is acceptable for veterinary use as well as human pharmaceutical use.

[0072] A “patient” is a human or non-human animal in need of medical treatment. Medical treatment can include treatment of an existing condition, such as a disease or disorder, prophylactic or preventative treatment, or diagnostic treatment. In some embodiments, the patient is a human patient.

[0073] A “therapeutically effective amount” of a pharmaceutical combination of this disclosure means an amount effective, when administered to a patient, to provide a therapeutic benefit such as weight loss or improvement in glycemic control.

[0074] The presently-disclosed subject matter is further illustrated by the following specific but non-limiting examples. The following examples may include compilations of data that are representative of data gathered at various times during the course of development and experimentation related to the presently disclosed subject matter. EXAMPLES EXAMPLE 1. Electrolyte complex (electrolyte complex) Synthesis

[0075] Cyanidin chloride (Reagent 1) was purchased from Chromadex chemicals. Sucrose octasulfate aluminum salt (Reagent 2) was obtained as previously described in USPN 11,524,024; USPN 10,716,802, and USPN 11,666,597.

[0076] Reagent 1 (1 part) was dissolved in water (10-2000 parts) to provide a dark purple solution. Reagent 2 (5 – 1000 parts) was added to the solution with mixing. After the addition of Reagent 2, the solution underwent a hypsochromic shift from dark purple to violet. The solution was stirred for 30 minutes until a uniform mixture was obtained. The suspension wasdried via open air, heated air drying, spray drying, and / or freeze-drying. The obtained solid was ground to provide the electrolyte complex. EXAMPLE 2. Berry Extraction Process

[0077] Frozen berries (D46-21, 1.567 kilograms, kg) were defrosted in water (1.5 liters, L) at 80ºC to provide a berry / water mixture. The berries comprised anthocyanin-rich berries and fruits, such as blueberries, bilberries, sloe berries, grapes or the like. Lafase Boost (0.036 milliliters, mL) were added to the berry / water mixture and the resultant mixture was stirred and mashed. The stirred and mashed mixture was left in a dark cabinet to macerate for 24 hours. After 24 hours, Lafase XL Press (0.15 mL) was added to the macerated mixture with stirring. Then the mixture was left standing for 24 hours. During standing the pH of the mixture was monitored, the pH at 2 hours (h) after the addition of Lafase XL press was 3.75, 4 h after the addition of Lafase XL press pH = 3.76, and 20 h after the addition of Lafase XL press pH = 4.04. Twenty-four hours after the addition of the Lafase XL Press, the mixture was heated to 88±3ºC and maintained for 15 minutes. The mixture was then allowed to cool with temperature monitoring. At 42ºC, the mixture was pressed through a sieve to remove the solid residue and provide 2060 mL of liquid filtrate. The filtrate was divided into two fractions (Fraction 1, 1010 mL and Fraction 2, 1050 mL). Salt (NaCl, 175 grams, g) was added to Fraction 2 and the solution was allowed to stand for 90 minutes as a precipitate formed. Fraction 1 and Fraction 2 were then filtered (pore size 25 – 30 μm). The polyphenol content in the fractions were 4.050 milligrams per mL (mg / mL) and 2.979 mg / mL, respectively. The fractions were purified by column chromatography (SepaBeads SP 70, 2 L of water, 70% ethanol desorption). Total elution of 1090 mL for Fraction 1 with 2.676 mg / mL of polyphenols. Total elution of 1030 mL for Fraction 2 with 2.296 mg / mL of polyphenols. The column extracts were concentrated in a rotary evaporator (425 mL Fraction 1 and 315 mL Fraction 2) and spray-dried to provide the product as a powder (5 g, 42.6% purity, Fraction 1 and 4 g, 41.3% purity Fraction 2). Details for product isolation of Fraction 1 and Fraction 2 are provided in Table 2 below. Table 2. Berry Extract Purification

[0078] The anthocyanin content of the isolated product was determined to constitute 3% wt (30 mg cyaniding equivalents per gram of sample). A representative FTIR spectrum of the isolated product is shown in FIG. 2. The qualitative composition of the extracts were analyzed by LC-MS and the identified components are provided in Table 3. Table 3. Chemical Composition of Berry ExtractEXAMPLE 3. Berry-Extract Based electrolyte complex Synthesis

[0079] Blueberry and sloe berry extracts with a total polyphenol content of >30% (Reagent 3) were used as to prepare electrolyte complexes. The synthesis of the electrolyte complexes was as described in EXAMPLE 1 with the exception that the Reagent 3 was used in place of Reagent 1. Changes in the molecular structure of the electrolyte complexes were confirmed by FTIR analysis as shown in FIG. 2. EXAMPLE 4. Glucose Tolerance Testing

[0080] Following a 12-hour fasting period, a portable continuous glucose monitoring device was used to monitor blood glucose levels over 120 minutes after ingestion of 75 grams of glucose as an aqueous solution. For the test subjects, one hour prior to glucose ingestion, the electrolyte complex of EXAMPLE 3 (650 mg), Reagent 2 (450 mg), or Reagent 3 (200 mg) were administered. As shown in FIG. 3, administration of the electrolyte complex resulted in the pronounced reduction in the blood glucose level. In comparison, the co- polymers (Reagent 2 and Reagent 3) did not demonstrate a comparable reduction in the blood glucose level. Some effect on blood glucose was observed during the test for Reagent 2, which prolonged the absorption of the glucose by extending the time for a blood glucose of <150 mg / dl.

[0081] Reagent 2 was previously tested by the group of Karp, et al. (USPN 11,524,024, USPN 10,716,802, USPN 11,666,597) in the rat model, whereby it demonstrated reduction in the absorption of glucose. The dose used by Karp was 500 mg / kg of body weight whereas the electrolyte complex used in the glucose tolerance testing was administered at a dose of 43 mg / kg of body weight.

[0082] A pronounced reduction in the absorption of glucose following administration of the electrolyte complex was observed with a 5-fold dose reduction as compared to previousstudies. Neither of the tested co-polymers provided a similar reduction in glucose absorption when administered in the quantity equivalent to the monomer quantity present in the electrolyte complex. This demonstrates that the electrolyte complex is distinctly superior to either of the co-polymers in reducing sugar absorption. EXAMPLE 5. Weight Loss Induction

[0083] The electrolyte complex or a vehicle control were administered to obese C57BL / 6 mice aged 9+ months with an average body weight of approximately 40 grams. For the test samples, 2.6 mg of electrolyte complex were suspended in 100 μl of water and administered two times per day for 14 consecutive days. For the control samples, 100 μl of water was administered as the vehicle control. Animals had ad libitum access to food and water during the experiment. Weight was measured before the start of the experiment and daily until the administration of the last dose. The weight loss results are shown in FIG. 4.

[0084] Oral administration of the EXAMPLE 3 electrolyte complex compound at a daily dose of 200 mg / kg of mouse body weight resulted in the weight loss of 13.26% over the course of 14 days. For comparison, Karp, et al. did not observe weight loss, but reported an 8% reduced weight gain when they administered LuCl compound to rats. Furthermore, the electrolyte complex dose corresponding to the LuCl dose used by Karp, et al. in rats would be 1000 mg / kg or more, which is 5-fold greater than the dose of the electrolyte complex.

[0085] These data confirm that electrolyte complexes obtained via crosslinking sucrose sulfate anions with flavylium cations are advantageous to the non-crosslinked preparations of sucrose sulfate which were reported previously, as indicated by the observations of electrolyte complex reducing glucose absorption and inducing weight loss. Distinct advantages include increased efficacy, reduced dose and, consequently, lower exposure to the compound, leading to reduced risks of toxicity and improved safety. EXAMPLE 6. Electrolyte complex Syntheses with Xanthan Gum

[0086] Berry extract as prepared in EXAMPLE 2 (2 parts) was dissolved in water (200 parts) and stirred until dissolved. Xanthan gum (1 part) was slowly added to the solution with mixing until a uniform suspension was formed. The suspension was dried. Suitable drying methods include drying in open air, drying in heated air, spray-drying, or freeze-drying. The solid was collected and ground to provide the Xanthan gum-based electrolyte complex. EXAMPLE 7. Electrolyte complex Synthesis with Sodium Alginate

[0087] Berry extract as prepared in EXAMPLE 2 (2 parts) was dissolved in water (200 parts) EXAMPLE 9. Weight Loss Induction with EXAMPLE 6 electrolyte complex

[0088] The EXAMPLE 6 electrolyte complex compound or a vehicle control were orally administered to obese C57BL / 6 mice aged 9+ months with an average body weight of approximately 40 grams. For the test samples, 2.6 mg of the EXAMPLE 6 electrolyte complex compound were suspended in 100 μl of water and administered two times per day for 14 consecutive days. For the control samples, 100 μl of water was administered as the vehicle control. For both test samples and controls the material was gavaged to the animals. Animals had ad libitum access to food and water during the experiment. Weight was measured before the start of the experiment and daily until the administration of the last dose. The weight loss results are shown in FIG. 6. The mice consuming the EXAMPLE 6 electrolyte complex compound lost an average of 9.2% of their initial body weight over the course of the two-week treatment. As indicated by the vehicle control data, a small percentage of the weight loss could be attributed to the experimental conditions, i.e. stress from the repeated gavage.

Claims

CLAIMS What is claimed is:

1. An electrolyte complex derived from a reaction mixture comprising an anionic saccharide, or a pharmaceutically acceptable salt thereof; and a flavylium cation, a glycoside thereof, an acylated derivative thereof, a pharmaceutically acceptable salt thereof, or a combination thereof.

2. The electrolyte complex of claim 1, wherein the anionic saccharide comprises an anionic monosaccharide, an anionic disaccharide, an anionic polysaccharide, a combination thereof, or a pharmaceutically acceptable salt thereof.

3. The electrolyte complex of claim 1, wherein the anionic saccharide comprises an anionic mucopolysaccharide, an anionic glycosaminoglycan, an anionic mucilage, a combination thereof, or a pharmaceutically acceptable salt thereof.

4. The electrolyte complex of claim 1, wherein the anionic saccharide comprises an anionic polysaccharide, or a pharmaceutically acceptable salt thereof, and optionally, an anionic monosaccharide, an anionic disaccharide, a combination thereof, or a pharmaceutically acceptable salt thereof.

5. The electrolyte complex of claim 1, wherein the flavylium cation is derived from cyanidin, delphinidin, pelargonidin, peonidin, petunidin, malvidin, rosinidin, luteolinidin, apigeninidin, aurantinidin, hirsutidin, tectoridin, capensinidin, a glycoside thereof, a pharmaceutically acceptable salt thereof, or a combination thereof.

6. The electrolyte complex of claim 1, wherein the flavylium cation comprises a glycoside.

7. The electrolyte complex of claim 6, wherein the glycoside comprises glucose, galactose, rhamnose, arabinose, xylose, glucuronic acid, sophorose, sambubiose, rutinoside, neohesperidose, gentiobiose, or a combination thereof.

8. The electrolyte complex of claim 1, wherein the ratio of the anionic saccharide to the flavylium cation in the reaction mixture ranges from 1:1 to 1000:

1.

9. The electrolyte complex of claim 1, wherein the ratio of the anionic saccharide to the flavylium cation in the reaction mixture ranges from 5:1 to 500:

1.

10. The electrolyte complex of claim 1, wherein the flavylium cation is derived from anthocyanin, and optionally, flavonol, coumarin, phenolic acid, or a combination thereof.

11. The electrolyte complex of claim 1, wherein the flavylium cation is derived from a berry extract.

12. The electrolyte complex of claim 11, wherein the berry extract is a polyphenol- enriched berry extract comprising at least 20 wt% total polyphenol content, as determined using a colorimetric assay.

13. The electrolyte complex of claim 12, wherein the polyphenol-enriched berry extract comprises 35-100 wt% polyphenol, preferably at least 70 wt%, more preferably at least 80 wt% polyphenol, based on the total dry weight of the extract.

14. The electrolyte complex of claim 1, wherein the anionic saccharide comprises a xanthan gum, an alginate, a sucrose sulfate, a pharmaceutically acceptable salt thereof, or a combination thereof.

15. The electrolyte complex of claim 1, wherein the anionic saccharide comprises xanthan gum, an alginate, a sucrose sulfate, a pharmaceutically acceptable salt thereof, or a combination thereof; and the flavylium cation is derived from an anthocyanin, a glycoside thereof, an acylated derivative thereof, a pharmaceutically acceptable salt thereof, or a combination thereof.

16. The electrolyte complex of claim 1, wherein the anionic saccharide comprises xanthan gum, an alginate, a sucrose sulfate, a pharmaceutically acceptable salt thereof, or a combination thereof; and the flavylium cation is derived from a berry extract.

17. The electrolyte complex of claim 16, wherein the berry extract is a polyphenol- enriched berry extract comprising at least 20 wt% polyphenol, based on the total dry weight of the extract.

18. The electrolyte complex of claim 17, wherein the polyphenol-enriched berry extract comprises 35-100 wt% polyphenol, preferably at least 70 wt%, more preferably at least 80 wt% polyphenol.

19. The electrolyte complex of claim 1, wherein the anionic saccharide is an alginate.

20. The electrolyte complex of claim 1, wherein the anionic saccharide is an alginate, and the flavylium cation is derived from an anthocyanin, a glycoside thereof, an acylated derivative thereof, a pharmaceutically acceptable salt thereof, or a combination thereof.

21. The electrolyte complex of claim 1, wherein the anionic saccharide is an alginate, and the flavylium cation is derived from a berry extract.

22. The electrolyte complex of claim 21, wherein the berry extract is a polyphenol- enriched berry extract comprising at least 20 wt% polyphenol, based on the total dry weight of the extract.

23. The electrolyte complex of claim 22, wherein the polyphenol-enriched berry extract comprises 35-100 wt% polyphenol, preferably at least 70 wt%, more preferably at least 80 wt% polyphenol.

24. The electrolyte complex of claim 1, wherein the anionic saccharide comprises a sucrose sulfate.

25. The electrolyte complex of claim 1, wherein the anionic saccharide comprises a sucrose sulfate, or a pharmaceutically acceptable salt thereof; and the flavylium cation is derived from an anthocyanin, a glycoside thereof, an acylated derivative thereof, a pharmaceutically acceptable salt thereof, or a combination thereof.

26. The electrolyte complex of claim 1, wherein the anionic saccharide comprises a sucrose sulfate, or a pharmaceutically acceptable salt thereof; and the flavylium cation is derived from a berry extract.

27. The electrolyte complex of claim 26, wherein the berry extract is a polyphenol- enriched berry extract comprising at least 20 wt% polyphenol, based on the total dry weight of the extract.

28. The electrolyte complex of claim 27, wherein the polyphenol-enriched berry extract comprises 35-100 wt% polyphenol, preferably at least 70 wt%, more preferably at least 80 wt% polyphenol, based on the total dry weight of the extract.

29. A pharmaceutical composition comprising the electrolyte complex of claim 1.

30. A pharmaceutical composition comprising a therapeutically effective amount of an active ingredient and excipients comprising the electrolyte complex of claim 1.

31. A composition comprising a polyphenol or a polyphenol-enriched extract and an additive comprising the electrolyte complex of claim 1.

32. A method for treating insulin resistance, gastrointestinal bloating, flatulence, dyslipidemia, polycystic ovarian syndrome, fatty liver disease, irritable bowel syndrome, and small intestinal bacterial overgrowth comprising administering to the patient in need thereof a therapeutically effective amount of electrolyte complex of claim 1.

33. A method for treating insulin resistance, gastrointestinal bloating, flatulence, dyslipidemia, polycystic ovarian syndrome, fatty liver disease, irritable bowel syndrome, and small intestinal bacterial overgrowth comprising administering to the patient in need thereof a therapeutically effective amount of the pharmaceutical composition of claim 29.

34. A method for enhancing athletic performance comprising administering to the patient in need thereof a therapeutically effective amount of electrolyte complex of claim 1.

35. A method for enhancing athletic performance comprising administering to the patient in need thereof a therapeutically effective amount of the pharmaceutical composition of claim 29.

36. method for treating or preventing obesity in a patient in need thereof, comprising administering to the patient in need thereof a therapeutically effective amount of electrolyte complex of claim 1.

37. A method for treating or preventing obesity in a patient in need thereof, comprising administering to the patient in need thereof a therapeutically effective amount of the pharmaceutical composition of claim 29.

38. A method for improving glycemic control in a patient in need thereof, comprising administering to the patient in need thereof a therapeutically effective amount of the electrolyte complex of claim 1.

39. A method for improving glycemic control in a patient in need thereof, comprising administering to the patient in need thereof a therapeutically effective amount of the pharmaceutical composition of claim 29.

40. A polyphenol-enriched berry extract comprising at least 20 wt% polyphenol, based on the total dry weight of the extract.

41. The polyphenol-enriched berry extract of claim 40 comprising 35-100 wt% polyphenol, preferably at least 70 wt%, more preferably at least 80 wt% polyphenol, based on the total dry weight of the extract.

42. The polyphenol-enriched berry extract of claim 40 free of organic solvents.

43. The polyphenol-enriched berry extract of claim 40 derived from blueberries, bilberries, blackberries, elderberries, black raspberries, chokeberries, cranberries, acai berries, sloe berries, mulberries, or a combination thereof.

44. A pharmaceutical composition comprising the polyphenol-enriched berry extract of claim 40 and a pharmaceutically acceptable excipient.

45. A method for preparing the polyphenol-enriched berry extract of claim 40 comprising macerating the berries in an aqueous solution in the presence of enzymes.

46. A method for treating or preventing obesity in a patient in need thereof, comprising administering to the patient in need thereof a therapeutically effective amount of electrolyte complex of claim 40.

47. A method for treating or preventing obesity in a patient in need thereof, comprising administering to the patient in need thereof a therapeutically effective amount of the pharmaceutical composition of claim 44.

48. A method for treating insulin resistance, gastrointestinal bloating, flatulence, dyslipidemia, polycystic ovarian syndrome, fatty liver disease, irritable bowel syndrome, and small intestinal bacterial overgrowth comprising administering to the patient in need thereof a therapeutically effective amount of electrolyte complex of claim 40.

49. A method for treating insulin resistance, gastrointestinal bloating, flatulence, dyslipidemia, polycystic ovarian syndrome, fatty liver disease, irritable bowel syndrome, and small intestinal bacterial overgrowth comprising administering to the patient in need thereof a therapeutically effective amount of the pharmaceutical composition of claim 44.

50. A method for improving glycemic control in a patient in need thereof, comprising administering to the patient in need thereof a therapeutically effective amount of the electrolyte complex of claim 40.

51. A method for improving glycemic control in a patient in need thereof, comprising administering to the patient in need thereof a therapeutically effective amount of the pharmaceutical composition of claim 44.

52. A method for enhancing athletic performance comprising administering to the patient in need thereof a therapeutically effective amount of electrolyte complex of claim 40.

53. A method for enhancing athletic performance comprising administering to the patient in need thereof a therapeutically effective amount of the pharmaceutical composition of claim 44.

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