JP2019178154A - Cell hydration composition comprising cyclodextrin - Google Patents

Abstract

To provide a composition that interacts with a biological cell line, comprising a biologically active molecule as well as a biomolecule surface, a cell component, and a water molecule having a specific density.SOLUTION: According to the present invention, a composition comprises a biologically active component constructed to increase the activity of a biological cell line by increasing the hydration of one or more components of the cell line. The biologically active component may comprise a first carbohydrate clathrate subcomponent, which increases the hydrogen bond structure of water, and a second solute subcomponent. The biologically active component may comprise an inclusion complex composed of a clathrate component and a complexing compound. The clathrate subcomponent may comprise amylose or cyclodextrin. According to the present invention, there are also provided a beverage and method for improving the hydration of cells in an animal, such as a human.SELECTED DRAWING: Figure 11

Description

The present invention relates generally to the regulation of biological cell activity, particularly cell activity that depends on the state of hydration. More particularly, the invention relates to biologically active components that are constructed to increase the activity of a biological cell line by increasing the hydration of one or more components of that cell line. The biologically active component may include a first carbohydrate clathrate subcomponent that increases the hydrogen bonding structure of water. The invention further relates to delivering biological compounds in vivo to alter the physiological activity of a mammal.

Water molecules interact primarily through hydrogen (H) bonds and through alignment of dipole moments. For example, the bond between adjacent water molecules is reinforced or stabilized by the alignment of the bond axis and the next adjacent water molecule. In liquid water, such an alignment propagates to the surrounding aqueous medium and a submicrometer scale molecular structure is established.

Examples of products and methods of use of cyclodextrins as clathrates to form inclusion bodies with bioactive guest molecules to improve the solubility and / or bioavailability of pharmaceutical compounds are Nos. 7,115,586 and 7,202,233, and US Patent Application Publication Nos. 2004/0137625 and 2009/0227690, the disclosures of which are incorporated herein by reference for all purposes.

Examples of products containing clathrates that bind to hydrophobic biomolecules and methods of use are described in US Pat. Nos. 6,890,549, 7,105,195, 7,166,575, the complete disclosures of which are incorporated herein by reference for all purposes. No. 7,423,027, and 7,547,459; US Patent Application Publication Nos. 2004/0161526, 2007/0116837, 2008/0299166, and 2009/0023682; Japanese Patent Application JP 60 -094912; Suzuki and Sato, `` Nutritional significance of cyclodextrins: indigestibility and hypolipemic effect of α-cyclodextrin '' J. Nutr. Sci. Vitaminol. (Tokyo 1985; 31: 209-223); and Szejtl et al., Staerke / Starch, 27 (11), 1975, pages 368-376.

US Patent Application Publication No. 2009/0110746 is a chemical agent that has the property of increasing the water diffusivity of dissolved molecular oxygen (O ₂ ) in the human body, and makes cyclodextrin the first oxygenation promotion. Can be included as a second “carrier” component to improve the solubility of the pro-oxygenating agent, and the cyclodextrin can enhance water diffusivity, tissue oxygenation, water structure, or cellular hydration Chemical agents that are not intended as direct altering agents are described.

U.S. Patent No. 7,115,586 U.S. Patent No. 7,202,233 US Patent Application Publication No. 2004/0137625 US Patent Application Publication No. 2009/0227690 U.S. Patent No. 6,890,549 U.S. Patent No. 7,105,195 U.S. Patent No. 7,166,575 U.S. Patent No. 7,423,027 U.S. Patent No. 7,547,459 US Patent Application Publication No. 2004/0161526 US Patent Application Publication No. 2007/0116837 US Patent Application Publication No. 2008/0299166 US Patent Application Publication No. 2009/0023682 Japanese patent application JP 60-094912 US Patent Application Publication No. 2009/0110746

Suzuki and Sato, “Nutritional significance of cyclodextrins: indigestibility and hypolipemic effect of α-cyclodextrin” J. Nutr. Sci. Vitaminol. (Tokyo 1985; 31: 209-223) Szejtl et al., Staerke / Starch, 27 (11), 1975, pp. 368-376 Hinrichs, W. et al., "An Amylose Antiparallel Double Helix at Atomic Resolution", Science, (1987), 238 (4824): 205-208. Lindner and Saenger (Carbohydr.Res., 99: 103, 1982) Segtan et al. (Anal. Chem. 2001; 73, 3153-3161) R. Giangiacomo (Food Chemistry, 2006, 96.3. 371-379)

A composition that interacts with a biological cell system comprising a biologically active molecule together with a biological molecule surface, a cellular component and a water molecule having a specific density, wherein the activity of said biological cell-based system is one or more A composition comprising a biologically active ingredient constructed to increase by increasing the hydration of the ingredient.

It is a figure which shows the chemical bond model of (beta) -cyclodextrin which is a cyclic oligosaccharide which has seven (alpha) [1-4] coupling | bonding-type glucose units. It is a figure which shows the structural model of cyclodextrin which has a toroidal topology entirely. FIG. 3 shows a structural model of cyclodextrin including the arrangement of glucosyl hydroxyl groups along the edge of the toroid. FIG. 3 shows the calculated dynamic distribution of molecules of water molecules around a β-cyclodextrin molecule 1 picosecond after the first contact. FIG. 5 shows the calculated molecular dynamic distribution of the water molecules of FIG. 4 including a more organized open water structure 1000 picoseconds after the first contact. FIG. 6 is a view showing an image subjected to threshold processing of the distribution of water molecules shown in FIGS. It is a figure which shows the comparison of the NIR differential spectrum including a specific wavelength range about the water sample in which cyclodextrin was melt | dissolved, and the water sample in which cyclodextrin is not melt | dissolved. It is a figure which shows the comparison of the NIR differential spectrum including a specific wavelength range about the water sample in which cyclodextrin was melt | dissolved, and the water sample in which cyclodextrin is not melt | dissolved. It is a figure which shows the comparison of the NIR differential spectrum including a specific wavelength range about the water sample in which cyclodextrin was melt | dissolved, and the water sample in which cyclodextrin is not melt | dissolved. It is a figure which shows the comparison of the NIR differential spectrum including a specific wavelength range about the water sample in which cyclodextrin was melt | dissolved, and the water sample in which cyclodextrin is not melt | dissolved. FIG. 4 shows a comparison of seed germination kinetics in water variably including cyclodextrin, amino acids, and cyclodextrin / amino acid inclusion complexes. FIG. 6 shows a comparison of seed germination kinetics in water variably including cyclodextrin, vitamins, and cyclodextrin / vitamin inclusion complexes. FIG. 3 is a diagram showing a comparison of seed germination rates in water variably including hydration active ingredients according to the present disclosure. FIG. 3 shows a comparison of long life of linear animals in a medium variably including cyclodextrin as an active component of hydration according to the present disclosure. 2 shows a comparison of the long life of linear animals in media variably including derivatized cyclodextrin as an active component of hydration according to the present disclosure. FIG. 3 is a diagram showing a comparison of long life of linear animals in a medium variably including a cyclodextrin inclusion complex as an active component of hydration according to the present disclosure. FIG. 2 is a diagram showing the death frequency of linear animals in media containing and not containing cyclodextrin inclusion complex as an active component of hydration according to the present disclosure. FIG. 4 shows population survival curves for linear animals living in media with and without cyclodextrin inclusion complex as active components of hydration according to the present disclosure.

The water structure is deliberately increased or organized by adding one or more solutes or appropriate molecular aggregates whose surface can compete strongly with water molecules for hydrogen bonding and / or dipole orientation. . In particular, factors and agents that enhance water molecule interactions and increase water structure thereby alter hydration or solvation of additional molecular surfaces. Thus, due to the first solution additive that increases the water structure, depending on the hydrogen bonding surface properties of the second component, the hydration interaction with the molecular surface of the second solution component (e.g., bond strength and kinetics) Can be increased, or such interactions can be decreased.

Furthermore, factors that alter the water structure generally change the average distance between water molecules, thereby increasing or decreasing the density of water. For example, when the temperature of the water drops below its freezing point, the hydrogen bonds between the water molecules overcome the kinetic energy of the water molecules, resulting in an increase in water structure and thereby a density of frozen water of approximately 9 %Decrease. Similarly, in liquid water, as the hydrogen bond strength of water increases, the average distance between water molecules increases, which is observed as an increase in specific volume (ie, a decrease in density). Decreasing the density of liquid water can increase the diffusivity of dissolved solutes. Therefore, the diffusion rate of co-dissolved solutes can be increased by aqueous additive components that reduce the density of water.

A chaotrope, as used herein, is a water-containing additive that functions to break the hydrogen bonding network in aqueous solution, thereby reducing the water structure. Chaotropes are generally less polar than water molecules and have a weak hydrogen bonding potential. Chaotropes can preferentially bind to nonpolar solutes and particles, thereby increasing the solubility of the nonpolar solutes.

Cosmotrope, as used herein, promotes a strong and extended hydrogen bonding network in aqueous solution, thereby increasing the sub-micrometer scale structure of water molecule interactions, and / or Or a stabilizing solute. A cosmotrope having a hydrogen bond chemical potential that exceeds the hydrogen bond chemical potential of water and / or a dipole moment that exceeds the dipole moment of water can increase the hydrogen bond network between water molecules. In addition, the cosmotrope can increase hydration interactions at the molecular surface that can include intermolecular binding sites by strengthening the hydration structure. Thus, the cosmotrope can be used as an aqueous solution additive that stabilizes molecular interactions.

Furthermore, the cosmotrope can increase the effective chemical activity of the dissolved co-solute. As the strength of hydrogen bonding interactions between water molecules increases, water becomes more open and conforms to a lower specific density and a higher specific volume. Thus, the addition of a cosmotrope to an aqueous solution can increase the diffusivity of one or more of the dissolved cosolute species or compounds by causing a decrease in density. Increasing the diffusivity of a solute species or compound can increase its reactivity, chemical potential, effective concentration, and availability.

As discussed herein, the clathrate component is an amphiphilic carbohydrate compound that is hydrophilic and has an outer surface that is strongly hydrogen bonded to water and an inner surface that is less hydrophilic. The inner surface of the clathrate can selectively bind to a molecular structure that is relatively non-polar or less hydrophilic than water.

An inclusion complex, as used herein, is a chemical complex formed between two or more compounds, where the first compound (also referred to as the host) is the second compound (guest (Also referred to as molecules) has a structure that defines a partially enclosed space that fits and binds to the first compound. A host molecule can be referred to as a clathrate and can reversibly or irreversibly bind to a guest molecule.

A biological cell, as used herein, is a functional metabolic unit that self-replicates a living organism and can survive as a unicellular organism or as a subunit of a multicellular organism, , Including lipid membrane structures containing a functional network of interacting biomolecules such as nucleic acids and sugars. Biological cells include prokaryotic cells, eukaryotic cells, and cells dissociated from multicellular organisms that may include cultured cells previously obtained from multicellular organisms.

A biological cell system, as used herein, is a network in which biological cells and / or intracellular elements are functionally interconnected, living cells, non-viable cells, organelles, and / or living organisms. It may contain molecules.

A bioactive molecule, as used herein, is a molecular compound that has a functional activity in a biological cell system.

A biomolecule, as used herein, is a molecular compound synthesized by a biological cell. Biomolecules include compounds that are typically synthesized by cells, and compounds that are synthesized by genetically engineered cells and chemically synthesized copies of compounds derived from cells.

A biomolecule surface, as used herein, is a boundary of atoms outside the biomolecule and may include biochemical interaction surfaces such as binding sites.

A cellular component, as used herein, is a functional element of a biological cell, including biomolecules, biomolecule complexes, organelles, polymer structures, membranes and membrane-bound structures, It may further include functional pathways and / or networks such as serial.

The density of a substance is the mass per unit volume of that substance under specific conditions of temperature and pressure.

The specific volume of the substance is a volume per unit mass of the substance, and can be expressed as, for example, m ³ / kg. The specific volume of a material is equal to the reciprocal of its density.

A biologically active ingredient, as used herein, is a molecular substance that modifies (increases or decreases) the activity of a biological cell system.

A bioactive agent, as used herein, is a substance that, when added to a biological cell system, or cellular component, causes a change in the biological activity of the system, or component thereof.

A water bonding structure, as used herein, refers to a network of hydrogen bonds that maintain and organize the orientation of water molecules in the liquid and solid states. As used herein, the water structure increases when hydrogen bonds between water molecules at a given temperature are strengthened and decreases when hydrogen bonds between water molecules at a given temperature are weakened.

Interaction between cellular components, as used herein, refers to a chemical bond between biomolecule surfaces. Such an interaction may involve binding between two biomolecules such as a ligand and its specific receptor. Alternatively, such interactions may involve binding between biomolecules and organelles such as cell membranes.

An extracellular signal, as used herein, is a biomolecule that can modify (increase or decrease) the activity of a cell when applied outside the cell. Extracellular signals can bind to components of the cell membrane (outer membrane) of the cell, or can cross the cell membrane and regulate intracellular activity. Extracellular signals can include, but are not limited to, extracellular matrix components; cell membrane components such as glycoproteins and glycolipids; antigens; and diffusible biomolecules such as nitric oxide.

Intracellular messenger, as used herein, is an internal component of a biological cell that has an active state and functions in an active state as an intermediate signal that transmits an extracellular signal to an intracellular target. .

A pharmacological agent, as used herein, is a synthetic chemical that binds to a biomolecule or biomolecule complex, thereby changing its activity.

The present invention includes active compositions that increase the activity of a cell line by increasing the hydration of one or more components of the biological cell line.

The active composition for modifying cell hydration preferably comprises a first carbohydrate clathrate component that increases the hydrogen bonding structure of water. In some examples, the active composition preferably includes a first carbohydrate clathrate component that increases the hydrogen bonding structure of water, and a second solute compound that may be a bioactive agent. In some examples, the active composition preferably includes an inclusion complex formed between the clathrate component and a complexing compound that may be a bioactive agent.

Biological cells are multi-compartment structures, including chemically active water-based chambers and lipid-based membranes. Cell structure and activity are highly selective between their biomolecular components, such as lipids, structural proteins, enzyme proteins, carbohydrates, salts, nucleotides, and other metabolic and signaling biomolecules. Derived from chemical bonding. The strength and specificity of biomolecule binding reflects a complementary chemical topology at the binding interface. Hydrophilic and / or hydrophobic surfaces generally dominate the chemical topology of the biomolecule binding interface. In aqueous systems, hydrophobic and hydrophilic interactions are substantially driven by competing hydration interactions with water molecules with concentrations above 50M.

Cell hydration, as used herein, refers to the interaction between water molecules and biomolecular components of a cell system. Cell hydration can be modified by changing the strength and / or kinetics of hydrogen bonding between water molecules and biomolecule surfaces.

Aqueous additives that modify the water structure can alter the strength, kinetics, and / or specificity of binding between cellular components by modifying the hydration of the binding surface of the biomolecule. For example, a water additive that increases the water structure can enhance the strength, kinetics, binding strength between the secreted intercellular signaling factor and the cognate receptor located in the plasma membrane of the potential target cell of that factor. And / or the specificity can be changed, thus biasing the results of the cell's signaling network (biasing the results).

Suitable clathrates as active components of cell hydration according to the present invention include amylose and cyclodextrins. Amylose is a linear polysaccharide with D-glucose units. As shown in FIG. 1, cyclodextrins are macrocyclic oligosaccharides of D-glucose units linked by α (1-4) glucose-to-glucose bonds. Amylose and cyclodextrins are easily prepared in large quantities from hydrolyzed starch. The preparation of cyclodextrins involves enzymatic conversion, most commonly using the enzyme cyclodextrin-glycosyltransferase produced by Bacillus strains.

As shown in FIG. 2, cyclodextrins can vary depending on the number of glucose units contained in the ring. Cyclodextrin species include α-cyclodextrin (6 units), β-cyclodextrin (7 units), γ-cyclodextrin (8 units), and δ-cyclodextrin (9 units). Parental cyclodextrins, as used herein, are natural, non-chemically derivatized α-cyclodextrin, β-cyclodextrin, and γ-cyclodextrin, each of 18 (α- ), 21 (β-) and 24 (γ-), free, unmodified hydroxyl groups.

As schematically shown in FIG. 3, cyclodextrins generally have a toroidal topology that resembles a truncated cone or is half the shape of an open-ended cylinder. Thus, a cyclodextrin can be described as including an outer chemical surface that includes an outer surface and an edge of a cylinder, and an inner chemical surface that surrounds an inner cavity (inside the cylinder).

The external surface of cyclodextrin contains a high density of hydrophilic chemical groups that hydrogen bond with water. In particular, the hydroxyl groups of the parent α-cyclodextrin, β-cyclodextrin and γ-cyclodextrin structures are all concentrated at the end of the cyclodextrin cylinder. More specifically, the hydroxyl (—OH) chemical groups of cyclodextrins are located along the edges of the cylinder and their orientation is sterically restricted. The hydroxyl group at the C (6) position of glucose can be referred to as the first OH group and is oriented counterclockwise with respect to the narrow open end of the cyclodextrin cylinder. The hydroxyl group of C (2) at the glucose position can be referred to as the second hydroxyl group and is the angle in the clockwise direction relative to the wider open end of the cyclodextrin cylinder.

The high density, constrained orientation of the cyclodextrin hydroxyl groups creates particularly strong hydrogen bonding surfaces at both ends of the cyclodextrin cylinder. Physicochemical analysis and solvation modeling of cyclodextrins show that water molecules in close proximity to cyclodextrins are fixed in position and have low angular (rotational) mobility. Usefully, each type of cyclodextrin differs in the diameter of the cylinder and the number of hydroxyl groups, and also in the number and mobility of strongly bound water molecules.

The hydrogen bonding activity of the cyclodextrin compound can propagate to the surrounding aqueous medium. As shown in FIGS. 4 and 5, kinetic modeling of cyclodextrin molecules introduced into a defined population of water molecules at standard temperature and pressure allows nanosecond water reorganization throughout the volume. Is caused. Figure 4 shows the population distribution 1 picosecond (ps) after starting the mixed simulation, and Figure 5 shows the redistribution of the same population at 1000 ps (1 nanosecond), where the water molecules are more open. The structure is taken.

In some examples, cyclodextrins can function as an active component of cell hydration through cosmotrophic activity that increases the water binding structure, and by increasing hydrogen bonding between water molecules, The sum is altered, thereby changing the strength, kinetics, and / or specificity of binding between cellular components.

In some examples, cyclodextrins can function as an active component of cell hydration through cosmotrophic activity that increases the binding structure of water, with lower specific densities due to stronger hydrogen bonds between water molecules. It has an open water structure that has (ie, a higher specific volume) and also increases the diffusion rate of the bioactive molecule. Such examples may include soluble bioactive molecules such as enzymes, enzyme substrates, nutrients, metabolites, cytokines, neurotransmitters, hormones, extracellular signals, intracellular messengers, or pharmacological agents.

The active component of cellular hydration that increases the rate of diffusion in water can modulate one of many biological processes limited by the rate of change in the concentration of the bioactive component. For example, neurotransmitter clearance from the synaptic cleft is generally associated with passive catabolism of glutamate from excitatory synapses in the mammalian brain and active catabolism of acetylcholine by the diffusion-limited enzyme acetylcholinesterase at vertebrate neuromuscular synapses. Including diffusion. Similarly, the activity of electroexcitable cells such as muscle cells is generally coordinated by diffusion-controlled changes in the concentration of intracellular second messenger signal calcium.

Cyclodextrin cell hydration activity can be modified either by increasing or decreasing by forming an inclusion complex with the complexing agent. The internal surface of cyclodextrin lacks hydroxyl groups and is less hydrophilic than the surrounding aqueous environment, thereby preferentially binding to co-solute molecules with low hydrophilicity and hydrogen bonding potential.

Carbohydrate clathrate compositions that increase the hydrogen bonding structure of interstitial and intracellular fluids when ingested by animals include cell membrane surface hydration structures and biomolecular solvation, which serve healthy cell functions Cell hydration can be improved. By improving cell hydration, healthy cell function, for example, solutes, nutrients, waste products, cytokines, metabolites and other molecular actions that support cell function, differentiation, repair, growth, and survival It can be supported by increasing the import, export and / or diffusivity of the agent and by stabilizing the cell membranes of vulnerable tissues such as muscles and nerves.

In some examples, carbohydrate inclusion complexes ingested by animals can increase the hydrogen bonding structure of water, thereby improving cell hydration and / or diffusivity of cellular components. In some examples, carbohydrate inclusion complexes ingested by animals increase the hydrogen bonding structure of water, thereby improving cell hydration and / or diffusivity of cellular components (i.e., complex). It can be dissociated to release the cyclodextrin clathrate component (not formed). In some examples, carbohydrate inclusion complexes can increase water structure and improve cell hydration without dissociation. In some examples, the carbohydrate inclusion complex can be dissociated into a clathrate component to increase water structure and cellular hydration, and the complexing agent further increases the water structure and / or Other beneficial properties such as nutrition or flavor can be provided.

The carbohydrate clathrate composition of the present invention can be in a variety of forms, including forming into a solid powder, tablet, capsule, caplet, granule, pellet, wafer, powder, instant beverage powder, effervescent powder, or effervescent tablet. Can be provided. Some carbohydrate clathrate compositions can also be formed as or incorporated into a hydrous beverage or other food product. Such carbohydrate clathrate compositions remain reasonably stable during storage, and therefore the clathrate component dissociates from the complexing agent, reducing the cosmotropic activity of the complex, thereby It may be an inclusion complex that does not form a stronger complex with another compound that reduces its ability to improve cellular hydration.

The present disclosure also provides a method for improving cell hydration in animals such as humans. For example, some methods make beverages by dissolving an inclusion complex formed by (a) a carbohydrate clathrate component and a complexing agent that can dissociate from the carbohydrate clathrate component under physiological conditions. And (b) ingesting the beverage orally into the animal, where the carbohydrate clathrate component dissociates from the complexing agent and alters the strength, extent, and kinetics of the hydrogen-bonded water structures on the cell's biomolecule surface. Steps may be included.

I. Carbohydrate clathrate composition The carbohydrate clathrate component includes, but is not limited to, α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, methylated β-cyclodextrin, 2-hydroxypropylated β-cyclodextrin Water-soluble β-cyclodextrin polymer, partially acetylated α-cyclodextrin, β-cyclodextrin and γ-cyclodextrin, ethylated α-cyclodextrin, β-cyclodextrin and γ-cyclodextrin, Any suitable carbohydrate, including carboxyalkylated β-cyclodextrin, α-cyclodextrin, quaternary ammonium salts of β-cyclodextrin and γ-cyclodextrin, amylose (e.g., acetylated amylose), and mixtures thereof. Include .

In a preferred embodiment, the carbohydrate clathrate can be selected based on a cosmotrope activity that increases water structure alone or in combination with other solutes. Preferred cyclodextrin cosmotropes may include α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, 2-hydroxypropyl-cyclodextrin, carboxymethylated cyclodextrin, and quaternary ammonium cyclodextrin.

Cyclodextrin derivatives are alkylated derivatives, hydroxyalkylated derivatives, alkoxyalkylated derivatives, acetylated derivatives, quaternary ammonium salt derivatives, carboxyalkylated derivatives, maltosylated derivatives And glucosylated derivatives. The alkyl group of the cyclodextrin derivative may be linear or branched and the length of the main chain may be 1 to 3 carbons with a total of 1 to 6, preferably 1 to 3 carbon atoms. You can do it. Some non-limiting examples of cyclodextrin derivatives include methylated beta-cyclodextrin, 2-hydroxypropylated β-cyclodextrin, water-soluble beta-cyclodextrin polymer, partially acetylated α- Cyclodextrin, β-cyclodextrin, and / or γ-cyclodextrin, ethylated α-cyclodextrin, β-cyclodextrin, and / or γ-cyclodextrin, carboxyalkylated β-cyclodextrin, α-cyclodextrin , Β-cyclodextrin, and / or quaternary ammonium salts of γ-cyclodextrin, and mixtures of any combination of these derivatives together with or in combination with one or more cyclodextrins. Exemplary cyclodextrin mixtures include combinations of α-cyclodextrin, β-cyclodextrin, and / or γ-cyclodextrin, each in a weight ratio range of about 1: 1: 1 to 2: 2: 1. It's okay. Cyclodextrins include hydrated crystals, including, but not limited to, hydrated and / or amorphous forms of α-cyclodextrin, β-cyclodextrin, and / or γ-cyclodextrin, and mixtures thereof. And / or in amorphous form.

When the carbohydrate clathrate composition is solid, the cyclodextrin component is present in a concentration range of about 10-90% w / w, or about 15-70% w / w, or about 15-60% w / w. It's okay. Preferably, the cyclodextrin component may be present in a concentration range of about 10-50% w / w, or about 15-40% w / w. More preferably, the cyclodextrin component may be present in a concentration range of about 20-25% w / w.

When the carbohydrate clathrate composition is in the form of a water-containing beverage, the cyclodextrin component has a concentration of about 0.01-75% w / v, or about 0.05-50% w / v, or about 0.1-25% w / v May exist in a range. Preferably, the cyclodextrin component may be present in a concentration range of about 0.1-10% w / v. More preferably, the cyclodextrin component may be present in a concentration range of 0.1-5% w / v.

The carbohydrate clathrate composition preferably includes a clathrate capable of forming an inclusion complex with various complexing agents such as amino acids, vitamins, flavorants, odorants, colorants and the like. Non-exclusive examples of carbohydrate clathrate components that can be combined with complexing agents to form inclusions include α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, 2-hydroxypropyl-cyclohexane. Mention may be made of dextrin, carboxymethylated cyclodextrin, quaternary ammonium cyclodextrin, amylose, amylose derivatives, or any desired mixture thereof.

The cyclodextrin clathrate component can be further selected based on the desired binding characteristics with the selected complexing agent. Non-limiting examples of acceptable cyclodextrins can include commercially available and government-regulated forms of α-cyclodextrin, β-cyclodextrin and γ-cyclodextrin. The number of glucose units determines the internal dimensions of the cavity and its volume, and can also determine the selectivity in forming an inclusion complex with the guest molecule. The selected complexing agent, when bound to a host cyclodextrin or other host carbohydrate clathrate, can modify the physicochemical properties of the complexed host such that its kosmotropic activity is increased.

When the clathrate component is in the form of an amylose component, the amylose component is expressed as a degree of polymerization (DP) in the range of DP = 10-900, more preferably DP = 20-200, most preferably DP = 30-80. It may contain the glucose units represented. Amylose derivatives include, but are not limited to, acetylated amylose. Preferably, the amylose component may have a structure comprising α1,4-linked D-glucopyranose in a helix configuration that defines a central cavity for binding hydrophobic molecules. For example, the A-starch helix and B-starch helix of V-amylose may comprise parallel, left-handed double helices that define a central cavity. The helix of the amylose inclusion complex is composed of hydrophobic forces created by host-guest interactions, intermolecular hydrogen bonds between adjacent amylose glucoses, and intramolecular hydrogen bonds formed by adjacent turns in the helix. Can be stabilized by. This complete disclosure is referred to Hinrichs, W. et al., “An Amylose Antiparallel Double Helix at Atomic Resolution”, Science, (1987), 238 (4824): 205-208, which is incorporated herein by reference for all purposes. I want. Amylose clathrate component may be used to form inclusion complexes with low molecular weight complexing agents such as non-limiting examples of flavors, colorants, vitamins, amino acids, and / or amines. it can.

When the composition containing the amylose clathrate component is solid, the amylose component is preferably about 10-90% w / w, or about 15-70% w / w, or about 15-60% w / w May be present in a range of concentrations. More preferably, the amylose component may be present in a concentration range of about 10-50% w / w, or about 15-40% w / w. Most preferably, the amylose component may be present in a concentration range of about 20-25% w / w. When the composition containing the amylose clathrate component is in the form of a water-containing beverage, the amylose component is preferably about 0.1-75% w / v, or about 1-50% w / v, or about 1-25% It may be present in the w / v concentration range.

II. Complexing Agents In some examples, the clathrate compositions disclosed herein may optionally include one or more amino acids, vitamins, flavorings, odorants, and / or other nutrients. Components, as well as complexing agents that may include combinations or mixtures of these agents may be included. The carbohydrate clathrate composition may further comprise one or more carbon-forming ingredients for use in forming a beverage product.

Complexing agents can form strong complexes with clathrate components to increase kosmotropic activity and thereby affect cell hydration. Alternatively, these agents can weakly complex with the clathrate component so that it can dissociate from the clathrate component to increase the water structure with the free clathrate component.

Non-limiting examples of amino acids suitable for forming inclusion complexes with the carbohydrate clathrate compositions of the present disclosure include aspartic acid, arginine, glycine, glutamic acid, proline, threonine, theanine, cysteine, cystine, alanine, Valine, tyrosine, leucine, isoleucine, asparagine, serine, lysine, histidine, ornithine, methionine, carnitine, aminobutyric acid (alpha-isomer, beta-isomer, and gamma-isomer), glutamine, hydroxyproline, taurine, norvaline , Sarcosine, its salts, and mixtures thereof. Mixtures of these amino acids N- alkyl C ₁ -C ₃ derivatives and N- acylated C ₁ -C ₃ derivatives as well as any amino acid or derivatives thereof, are also encompassed. Preferred complexing amino acids that can be included in cyclodextrins to increase water structure and cellular hydration include L-arginine, L-lysine, N-methyllysine, and L-carnitine.

Non-limiting examples of vitamins, nicotinamide (vitamin B _3), niacinamide, niacin, hydrochloric pyridoxal (vitamin B _6), ascorbic acid, edible ascorbic acid esters, riboflavin, pyridoxine, thiamine, vitamin B _9, folic acid Folate, pteroyl-L-glutamic acid, pteroyl-L-glutamate, salts thereof, and mixtures thereof. Preferred vitamins that can be included in cyclodextrins to increase water structure and cellular hydration include nicotinamide and niacinamide.

Non-limiting examples of flavors include apple, apricot, banana, grape, black currant, raspberry, peach, pear, pineapple, plum, orange, and vanilla. Examples of flavoring related compounds include butyl acetate, butyl isovalerate, allyl butyrate, amyl valerate, ethyl acetate, ethyl valerate, amyl acetate, maltol, isoamyl acetate, ethyl maltol, isomaltol, diacetyl, ethyl propionate , Methyl anthranilate, methyl butyrate, pentyl butyrate, and pentyl pentanoate. Flavoring agent, the cyclodextrin component is selected from about 10～800M ^-1, preferably 30 to 150 m ^-1, selected to more preferably weaker bonds binding constant in the range of 40 to 100 M ^-1 be able to.

Non-limiting examples of other flavor improving ingredients include polyol additives such as erythritol, maltitol, mannitol, sorbitol, lactitol, xylitol, inositol, isomalt, propylene glycol, glycerol (glycerin), threitol, galactitol. , Palatinose, reduced isomaltooligosaccharide, reduced xylo-oligosaccharide, reduced gentio-oligosaccharide, reduced maltose syrup, and reduced glucose syrup.

Non-limiting examples of colorants include colorants that are known to have high water solubility and low lipophilicity. Examples of colorants having these properties are, for example, betalines such as betacyanine and betaxanthine, including bulgaxanthin, miraxanthin, portulaxanthin and indicaxanthin; anthocyanidins such as auranthidine, cyanidin, delphinidin , European pinidine, luteolinidine, pelargonidin, malvidin, peonidin, petunidin and rosinidine, and all the corresponding anthocyanins (or glucosides) of these anthocyanidins; and phenolic curcuminoids such as curcumin, demethoxycurcumin and bisdemethoxycurcumin Turmeric colorants included.

All examples of amino acids, vitamins, flavorings and related compounds described above may be in the appropriate salt or hydrated form.

The complexing agent can be selected to form an inclusion complex with the selected clathrate component. The complexing agent can bind to the clathrate component as a guest molecule in the cavity of the clathrate molecule and / or the selected weak complexing agent is clathrate at or near the clathrate edge. A so-called outer sphere complex bound to a molecule can be formed. For example, the selected weak complexing agent can be attached to the cyclodextrin molecule, to or around the primary hydroxyl group and / or the secondary hydroxyl group at the ring edge of the cyclodextrin. Some complexing agents that form outer sphere complexes with selected cyclodextrins are molecules that dissolve and form self-aggregating hydrated cyclodextrin molecules between two adjacent cyclodextrin molecules in water By shielding interhydrogen bonds, it can be reduced or prevented.

When the carbohydrate clathrate composition is solid, the complexing agent may be present in a concentration range of about 1-50% w / w. Preferably, the complexing agent may be present in a concentration range of about 1-40% w / w or about 1-25% w / w. More preferably, the complexing agent may be present in a concentration range of about 5-15% w / w.

When the carbohydrate clathrate composition is in the form of a water-containing beverage, the complexing agent may be present in a concentration range of about 0.1-25% w / v or about 1-20% w / v. Preferably, the complexing agent may be present in a concentration range of about 1-15% w / v or about 1-10% w / v or about 3-8% w / v. More preferably, the complexing agent may be present in a concentration range of about 5-8% w / v.

III. Inclusion Complex As described above, an inclusion complex may comprise a clathrate host molecule complexed with one or more complexing agents. In the form of a solid product such as a solid powder or tablet, the inclusion complex is essentially unique compared to a solid composition containing the same ingredients but without the formation of a preliminary inclusion complex. The nature of An inclusion complex is basically a chemical entity having a non-covalent hydrogen bond formed between a clathrate molecule and a weak complexing agent molecule. In the solid state, the clathrate is dissolved in a water-containing beverage, for example, when the clathrate component for increasing the water structure and the water structure are further increased, or when the inclusion complex is introduced into an aqueous environment. It has the potential to dissociate into complexing agents that can provide other beneficial properties such as nutrition or flavor, such as when ingested or ingested.

In the form of a solid product, the clathrate component and the one or more types of complexing agent may be substantially in the form of an inclusion complex as described above. Preferably, greater than about 25% of the clathrate component is complexed with one or more types of complexing agents in the form of an inclusion complex. In this order, over 35%, over 45%, over 50%, over 60%, over 70%, over 80%, over 90%, and over 95% of the clathrate components are complexed. More preferable.

IV. Carbonation Forming Components Some clathrate compositions may include a carbonation forming component that when carbonated or foamed when dissolved in an aqueous environment. The carbonic acid forming component can advantageously inhibit the self-aggregation of clathrate molecules, thereby structuring water and increasing the surface area of the clathrate to increase cell hydration.

Non-limiting examples of carbonic acid forming components can include sodium carbonate, sodium bicarbonate, potassium carbonate and potassium bicarbonate. Preferred carbonate forming components include sodium carbonate and sodium bicarbonate.

When the carbohydrate clathrate composition is solid, the carbonic acid forming component may be present in a concentration range of about 1-60% w / w or about 5-60% w / w. Preferably, the carbonic acid forming component may be present in a concentration range of about 5-45% w / w or 10-45% w / w. More preferably, the carbon-forming component may be present in a concentration range of about 10-15% w / w.

When the carbohydrate clathrate composition is in the form of a water-containing beverage, the carbon-forming component may be present in a concentration range of about 1-30% w / v or about 1-25% w / v. Preferably, the carbonic acid forming component may be present in a concentration range of about 2-15% w / v or 2-10% w / v. More preferably, the carbonogenic component may be present in a concentration range of about 2-5% w / v.

V. Other ingredients Some compositions may contain other ingredients that affect the flavor and / or nutritional value of the composition. These additional ingredients can include, but are not limited to, one or more of the following: serving as a clathrate anti-agglomeration additive in flavor additives, nutritional ingredients and / or formulations Various hydroxyl acids. Non-limiting examples of such other ingredients include citric acid, ascorbic acid, sodium chloride, potassium chloride, sodium sulfate, potassium citrate, europium chloride (EuCl ₃ ), gadolinium chloride (GdCl ₃ ), terbium chloride (TbCl ₃ ), magnesium sulfate, alum, magnesium chloride, maltodextrin, mono-, di-, tri-basic sodium or potassium salts of phosphate (e.g. inorganic phosphate), hydrochloride (e.g. inorganic chloride) And sodium hydrogen sulfate. Non-limiting examples of hydroxyl acids that prevent cyclodextrin aggregation can include isocitric acid, citric acid, tartaric acid, malic acid, threonic acid, salts thereof, and mixtures thereof. These hydroxyl acids can also exhibit several nutritional benefits. Other non-limiting examples of additional optional ingredients such as flavor additives that can be used include suitable organic salts such as choline chloride, alginic acid sodium salt (sodium alginate) , Glucoheptonic acid sodium salt, gluconic acid sodium salt (sodium gluconate), gluconic acid potassium salt (potassium gluconate), guanidine hydrochloride, glucosamine hydrochloride, amiloride hydrochloride, sodium glutamate (MSG) , Adenosine monophosphate, magnesium gluconate, potassium tartrate (monohydrate), sodium tartrate (dihydrate), and the like.

Preferred other components include, for example, citric acid, ascorbic acid, and maltodextrin.

When the carbohydrate clathrate composition is solid, the one or more other ingredients may be present in a concentration range of about 1-30% w / w or about 1-25% w / w, respectively. Preferably, the one or more other ingredients may be present in a concentration range of about 1-20% w / w or 1-15% w / w, respectively. More preferably, the one or more other components may each be present in a concentration range of about 2-5% w / w.

When the carbohydrate clathrate composition is in the form of a water-containing beverage, the one or more other ingredients may be present in a concentration range of about 1-20% w / v or about 1-15% w / v. Preferably, the one or more other components may be present in a concentration range of about 1-10% w / v or 1-5% w / v. More preferably, the one or more other components may be present in a concentration range of about 1-3% w / v.

VI. Ingredient ratios In addition to the above discussion regarding the types and amounts of various ingredients that can be used in the carbohydrate clathrate compositions disclosed herein, the same applies to the relative amounts of these ingredients. It is also noted that this can be explained. Preferably, the weight ratio of the clathrate component and complexing agent is from about 5: 1 to 1: 10 can be in the range of, more preferably, from about 2: 1 to 1: I in the range of 5 Der More preferably, it may be in the range of about 2: 1 to 1: 2, and even more preferably in the range of about 1: 1 to 1: 2.

With respect to other possible ingredients, such as flavor ingredients, carbonic acid forming ingredients, and other above-mentioned ingredients, the weight ratio of the clathrate ingredient to each other ingredient is independently about 25: 1 to It may be in the range of 1:25, or about 10: 1 to 1:10, or about 5: 1 to 1: 5, or optionally about 2: 1 to 1: 2, as well as 1: 1.

VII. Preferred Embodiments Preferred embodiments of the carbohydrate clathrate compositions disclosed herein are provided by way of example and are not intended to limit the scope of the present disclosure in any way.

Effect of cyclodextrin on molecular dynamics of water structure A simulated water solvated cyclodextrin molecular system was reported by Lindner and Saenger using HyperChem® 5.11 software (HyperCube Inc, Gainesville, FL) (See Carbohydr.Res., 99: 103, 1982) A total of 984 waters were obtained using input parameters obtained from single crystal analysis of cyclohepta-amylose dodecahydrate clathrate (or β-cyclodextrin). It was created using a periodic solvent box (3.1x3.1x3.1nm ³ ) of water containing molecules. Molecular transformations and atom types were adjusted using TinkerFFE 4.2 (TINKER Software Tools for Molecular Design, Version 5.0, Jay William Ponder, Washington University, St. Louis, MO) to the appropriate format. Molecular dynamics and dynamics calculations are performed after preliminary optimization using Tinker 5.0 software and the shortened Newton-Raphson method using Linux® x86-64 operating system (Slamd 64 v12.2) did.

Molecular dynamics simulations were performed using MM3 Force Field molecular mechanics software with steps of 120 picoseconds (psec) and 0.1 femtoseconds (fsec) at a constant temperature (298K). Records were generated by dumping the intermediate structure every 100,000 steps (equivalent to 10 psec elapsed time).

Observation results:
At zero time for each simulation, the standard water solvent box contained one β-cyclodextrin clathrate molecule and a population of 984 uniformly distributed water molecules. 4 and 5 show representative examples of the central portion of the solvent box at a particular elapsed time during one representative simulation. It will be appreciated that the position and orientation of the water molecules are shown as (bent) bars and β-cyclodextrin is shown as the van der Waals surface. Furthermore, it should be appreciated that FIGS. 4 and 5 show the volume of the solvent box, so that the three-dimensional molecular distribution is compressed in two dimensions.

FIG. 4 shows the central part of the solvent box at a simulation elapsed time of 1 psec. In particular, at an elapsed time of 1 psec, the water molecule immediately adjacent to β-cyclodextrin has an acquired relatively static (stable) position through hydrogen bonding with the cyclodextrin. Such water molecules can be referred to as the first hydration layer. However, the distribution of most water molecules in the solvent box generally remains similar to the initial unstructured distribution (before 1 psec).

FIG. 5 shows a simulation after an elapsed time of 1000 psec (ie, 1 nsec). At 1000 psec, water molecules immediately adjacent to β-cyclodextrin continue to occupy relatively static (stable) positions. However, compared to 1 psec (FIG. 4), the water molecules beyond the first hydrated layer acquired a more open microstructure.

Differences in water structure can be more easily observed when there is no explanation for the perspective shadows included in FIGS. FIG. 6 shows an alternative view of the distribution of water molecules shown in FIG. 4 (left side, labeled 1 psec) and FIG. 5 (right side, labeled 1000 psec), which was generated by the following method: 4 and 5 image files with 256 gradation levels (8 bits) are opened in Photoshop 9.0 (Adobe, Inc), adjusted to 300 dpi, thresholded at gradation level 207; image using circle selection tool Was trimmed so that the outer ring had the same diameter, the outer square corners were filled with black (tone level 0), and then further trimmed to blacken the inside of the ring with little cyclodextrin molecules. The outer and inner dimensions of the annulus were equally applied to the images being compared. The resulting thresholded display qualitatively shows that the water molecules around the central (closed) cyclodextrin molecule at 1000 psec have a more open and harmonized structure (e.g., right side of FIG. 6 panel).

In order to quantitatively evaluate the changes in water microstructure shown in FIGS. 4-6, the molecular density was estimated by measuring the open path through the indicated volume, which is the mean free path Similar to analysis, the mean free path in a defined volume of molecular material is inversely related to the density of the molecule. In particular, white pixel elements indicate open paths between water molecules, and the number of open paths in volume is easily quantified using the Photoshop 9.0 histogram tool for counting the number of white pixel elements. Applying to the panel of FIG. 6, the measured 2% increase was calculated for the open path at an elapsed time of 1000 psec compared to the open path at an elapsed time of 1 psec. For comparison, freezing pure water reduced the density by 9%. Since the path length is inversely proportional to the molecular density, analysis shows that dissolved cyclodextrin reduces the density of the aqueous solution by increasing the organization of water molecules.

In summary, the results show that after rapid (psec) hydrogen bonding adhesion between hydroxyl and water molecules on the outer surface of β-cyclodextrin, propagation of slower (nanosecond) water molecule reorientation throughout the solvent box Followed by a more open water structure. The measurement results further show that cyclodextrin can sufficiently increase the hydrogen bonds between water molecules around the water volume and reduce the density of water.

Effects of cyclodextrin additives on water binding detected by IR spectroscopy Physical microstructure tests on the detection of water, water-sugar interactions, and sugar effects on water structure increase and decrease, for example, Segtan (See Anal. Chem. 2001; 73, 3153-3161), and R. Giangiacomo (see Food Chemistry, 2006, 96.3. 371-379), in particular near-red Outer (NIR) spectroscopy was preferentially used.

Hydration binding energies in solutions of pure water and the same water containing cyclodextrin compounds were assayed using IR spectroscopy in the near infrared and mid infrared regions. To record a linear signal throughout the wavelength range, a short optical length cuvette was used to minimize water absorption attenuation.

The spectrum in the NIR region was registered with a FOSS NIR System, Inc. 6500 spectrometer and Sample Transport Module (STM) using a 1 mm-es cuvette. Transmission spectra were collected from 1100-2498 nm using a lead sulfide (PbS) detector and Vision 2.51 software (2001; FOSS NIRSystems, Inc.).

Using a Perkin-Elmer Spectrum 400 FT-NIR / FT-IR spectrometer and a UATR (Universal Attenuated Total Reflectance; ZnSe-diamond crystal, 1x flat plate) sample handling unit, spectra ranging from 2500 to 15385 nm were obtained. (Reported as 4000-650 cm ^-1 ). Measurements were performed at 24 ° C. using a triglycine sulfate (TGS) detector and Spectrum ES 6.3.2 software (PerkinElmer, 2008).

Three water samples were used in this study. The first water sample (USA I) was obtained from the United States and purified by reverse osmosis, carbon filtration, UV exposure, 0.2 micron, absolute filtration type membrane filtration, and ozone treatment. A second water sample (USA II) was obtained from the United States as well. A third water sample (BP I) was obtained from Budapest, Hungary. Capillary electrophoresis revealed similar ionic components with different concentrations among the three waters.

The following cyclodextrins were added to the above water sample in a concentration range of 0.1% to 5% w / v:
α-Cyclodextrin (also called αCD, ACD), Lot. No. CYL-2322.
β-cyclodextrin (also called βCD, BCD), Lot. No. CYL-2518 / 2.
γ-cyclodextrin (also called γCD, GCD), Lot. No. CYL-2323.
2-Hydroxypropyl-β-cyclodextrin (HPβCD, HPBCD), DS * = 3.5, Lot. No. CYL-2232.
2-Hydroxypropyl-γ-cyclodextrin (HPCD, HPGCD), DS * = 4.8, Lot. No. CYL-2258.
Carboxymethyl-β-cyclodextrin (CMBCD), Lot. No. CYL-2576.
Quaternary ammonium-β-cyclodextrin (QABCD).

In some examples, there is a variety between cyclodextrin and complexing bioactive agents including the amino acids L-arginine and L-carnitine and the vitamin niacinamide (also known as nicotinamide). The inclusion complex was formed. All reagents were analytically pure. In some examples, L-arginine and nicotinamide were formed in free form or in a cyclodextrin complex to assess the independent and co-dependent activities of cyclodextrin and bioactive agent ( Added in a molecularly confined form. The concentration of the additive in the above free form and in the form of cyclodextrin inclusion complex was in the range of 0.1% to 5.0% w / v.

Observation results:
FIG. 7 shows the second derivative NIR spectrum for the wavelength range 900 to 1200 nm. The results show that water-binding interactions are significantly modified by adding QABCD and are further significantly modified by adding CMBCD and HPBCD.

FIG. 8 shows the second derivative NIR spectrum shown for 1200-1500 nm. The results show that the water-binding interaction is significantly modified by adding QABCD and HPBCD, and is further significantly modified by adding CMBCD.

FIG. 9 shows the second derivative NIR spectrum shown for 1620-1710 nm. The results show that water-binding interactions are significantly altered by adding CMBCD, QABCD, and HPBCD.

FIG. 10 shows the second derivative NIR spectrum shown for 2170-2370 nm. The results show that the water-binding interaction is significantly modified by adding CMBCD and HPBCD, and more significantly by adding QABCD.

As shown in FIGS. 7-10, the addition of cyclodextrin changes the molecular binding interaction of the aqueous medium. In particular, with reference to FIG. 9, the refined NIR differential spectrum in the wavelength range of 1620-1770 nm shows the CH3-group, CH2-group and CH2 of the cyclodextrin additive to changes related to carbon-hydrogen bonds. -Indicates that a group is involved. Significant spectral changes that occurred in each of the cyclodextrin-treated water samples indicate that the cluster system in bulk water is dominated by the altered microstructure of hydrogen bonds. This effect was greatest in water samples treated with charged quaternary ammonium-β-cyclodextrin (QABCD), for example, as shown in FIGS.

Accelerated germination of plant embryos Wheat seeds (Triticum aestivum) were germinated using USA I water, USA II water, and BP I water as described in Example 2. Germination rates using unsupplemented (control) water were compared to germination rates using the same water with various supplements of cyclodextrin components and / or bioactive agents as active components of cell hydration. For each condition, 10 seeds were continuously contacted with water in a Petri dish and maintained at 25 ° C. with a 12 hour light / dark cycle. Photometric images were recorded from day 1 to day 6 after sowing. The percentage of germinated seeds was calculated and compared as a function of the percentage and time and the concentration of additive applied.

Water samples for germinating seeds alone, without additives, or containing cyclodextrin, or cyclodextrin and L-arginine or nicotinamide (both Sigma Chemical Co .; St. Louis, MO Or a clathrate inclusion complex with L-carnitine (Lonza AG; Switzerland). Additives were included at 0.1% (w / v) and 5.% (w / v). The additive solution was prepared fresh on the day of germination.

The parent cyclodextrins α-cyclodextrin (ACD), β-cyclodextrin (BCD), and γ-cyclodextrin (GCD) were obtained from Wacker Chemie (Munich, Germany). The following derivatized cyclodextrins were obtained from Cyclolab Ltd. (Budapest, Hungary): hydroxypropylated beta-cyclodextrin (DS-3) (HPBCD), carboxymethylated β-cyclodextrin (DS-3.5) (CMBCD), 2-hydroxy-3-N, N, N-trimethylamino) propyl-β-cyclodextrin chloride (DS˜3.6) (QABCD).

Observations Germination kinetics in control water and water modified with additives under the same conditions were quantified as a percentage of seeds with shoots. Each decision consisted of 100 seeds for each parameter. The results are reported in Table 1 below and Figures 11-13.

A) Cyclodextrin / L-Arg inclusion complex increases seed germination.

Table 1 shows 0.5% w / v α-CD, 0.5% w / v L-arginine (L-Arg), and 0.5% w / v α-CD / L- dissolved in USA I water, respectively. Figure 2 shows the comparative effect of arginine inclusion complex on wheat seed germination. The results in the table above show that the inclusion complex (αCD / L) between 0.5% (w / v) α-cyclodextrin and L-arginine compared to pure water lacking any additives (control). The wheat seed germination rate in water containing -Arg inclusion complex) is shown to be much higher. Furthermore, the results in Table 1 show that the seed germination rate in water containing an inclusion complex between α-cyclodextrin and L-arginine (αCD / L-Arg inclusion complex) is 0.5% (w / v) Compared with water containing α-cyclodextrin (αCD) alone as an additive and with water containing 0.5% (w / v) L-arginine (L-Arg) alone as an additive. It is much higher. Therefore, this result shows that the complex of α-cyclodextrin and L-arginine has a synergistic effect on the seed germination rate that is not shown by any of the individual components of the complex used as a single additive. Is shown. The results in Table 1 are also shown in FIGS. 11 and 13.

B) Cyclodextrin / nicotinamide inclusion complex increases seed germination.

Table 2 shows 0.5% w / v α-cyclodextrin, 0.5% w / v nicotinamide, and 0.5% w / v α-cyclodextrin / nicotinamide inclusion complex each dissolved in USA I water ( The comparative effect of αCD / nicotinamide inclusion complex) on the germination of wheat seeds is shown. The results in the above table show that the wheat seed germination rate is much higher in water containing inclusion complex between α-cyclodextrin and nicotinamide compared to pure water lacking any additives (control). Indicates. Furthermore, the results in Table 2 show that wheat seed germination rate in water containing inclusion complex between α-cyclodextrin and nicotinamide (αCD / nicotinamide inclusion complex) was added with α-cyclodextrin (αCD). Compared to water containing alone as a product and to water containing nicotinamide alone as an additive, it is much higher. Thus, the results show that α-cyclodextrin and nicotinamide have a synergistic biological activity that significantly increases seed germination rate when used as an inclusion complex. Such biological activity was not demonstrated by any of the individual components of the complex used as a single additive. The results in Table 2 are also shown in FIGS. 12 and 13.

C) Using USA II water and BP I water for germination, qualitatively similar results were obtained as reported in Tables 1 and 2 and Figures 11-13. Thus, in particular, cyclodextrin inclusion complexes containing L-arginine or nicotinamide are shown above using USA I water when dissolved in USA II water or BP I water, respectively. As shown, the wheat seed germination rate was significantly increased.

D) The length of sprout (rate of sprout growth during germination) did not differ between conditions with statistically significant confidence intervals (P <0.05). This result indicates that cyclodextrins, particularly cyclodextrin inclusion complexes, are selectively used as active components of cell hydration to promote seed germination without necessarily affecting the growth rate of shoots. It shows that it can be used.

C. elegans extended life of nematode (C. elegans) in water with modified hydration using USA I water (control) lacking any other additive ingredients (described in Example 2) Or alternatively, prepared using the same water supplemented with the parent α-cyclodextrin, β-cyclodextrin, or γ-cyclodextrin, and / or bioactive agent as the active component of cell hydration Grown in petri dishes containing liquid nutrient media. 50 ± 3 worms were transferred to each dish. Each condition was repeated in triplicate. The experiment was repeated for USA II water and BP I water as described in Example 2.

Additive to water:
A. Add parent α-cyclodextrin, β-cyclodextrin and γ-cyclodextrin.
B. Add L-arginine and nicotinamide.
C. Addition of cyclodextrin and inclusion complex of L-arginine and nicotinamide.

Observation results:
The recorded results are shown in Tables 3-5 below and are further shown in FIGS.

Table 3 shows middle-aged (10 days), elderly (15 days) and middle-aged (10 days) and medium in variably containing parent α-cyclodextrin, β-cyclodextrin, and γ-cyclodextrin as active components of cell hydration. The percentage of animals that survive to old age (18 days) is reported. In this example, the parent cyclodextrin was added to a nutrient medium dissolved in USA I water at a concentration of 0.1% w / v.

Consistent with all of the previous tests, in this example, normal nematode animals survived in normal medium for 2 weeks. Each of the parent cyclodextrins significantly increased the survival (percentage of survival) of the nematode at the age of long life (10-15 days). Furthermore, α-cyclodextrin and γ-cyclodextrin significantly increased the number of animals surviving until old age, ie after 15 days. The results are also shown graphically in FIG. 14, which compares the cumulative percentage of animals that survive to 15 and 18 days in media containing each of the parent cyclodextrins as an additive. The results show that parent cyclodextrins, in particular α-cyclodextrin and β-cyclodextrin, can be used as active components of cellular hydration to improve biological functions of living animals Yes.

Biological mechanisms that support aging may include a wide range of cellular activity improvements during aging, or by selectively activating a slowly aging cellular activity pathway. The clathrate-induced water structure, cellular component hydration, and increased diffusion of biologically active cellular components, including intercellular and intracellular signals, all contribute to the overall cyclodextrin's overall survival of the organism. Can contribute to the effect.

Table 4 shows middle-aged (10 days), old-aged (15 days) in medium variably containing derivatized α-cyclodextrin, β-cyclodextrin, and γ-cyclodextrin as active components of cell hydration. ) And the percentage of animals alive until old age (18 days). In this example, derivatized cyclodextrin was added to a nutrient medium dissolved in USA I water at 0.1% w / v.

As listed in Table 4, HP-derivatives, carboxymethyl-derivatives, and quaternary ammonium-derivatives of β-cyclodextrin have only a minor effect on the first 10-day survival of nematodes. did. In contrast, a significant increase in survival was observed at older age (15 days), but no significant increase was observed at older age (18 days). The results are also shown graphically in FIG. 15, comparing the cumulative percentage of animals surviving to 15 and 18 days in media containing each of the derivatized cyclodextrins as an additive. . The results show that derivatized cyclodextrins can be used as the active component of cellular hydration to improve the biological function of living animals.

Table 5 shows middle-aged (10 days), old-aged (14 days) in nutrient media dissolved in USA I water variably supplemented with cyclodextrin inclusion complex at 0.1% w / v as the active component of cell hydration. Days), and the percentage of animals that survive to old age (18 days). In this example, the inclusion complex contained α-cyclodextrin and a bioactive agent, specifically L-arginine, L-carnitine, or niacinamide.

As in the previous example, nematode animals survived for 2 weeks on unsupplemented medium. Α-Cyclodextrin complexes with L-arginine and niacinamide more than doubled the survival of nematodes in the elderly (day 14), and a small but significant number of animals survived on nutrient medium alone. It became possible to survive to old age when there was no animal that had been. In contrast, the α-cyclodextrin complex with L-carnitine had little or no significant effect on nematode survival. Similarly, L-arginine and nicotinamide added alone to the medium without α-cyclodextrin had little effect on nematode survival. The results are also shown graphically in FIG. 16, which compares the cumulative percentage of animals that survive to 14 and 18 days in media containing each cyclodextrin inclusion complex as an additive. The result shows that α-cyclodextrin inclusion complexes, especially complexes with L-arginine and niacinamide, can be used as active components of cellular hydration to improve the biological function of living animals. It shows what you can do.

As further shown in FIG. 17, the inclusion complex of α-cyclodextrin and L-arginine (data series A; 1: 1 complex, dissolved in 0.1% w / v in a medium prepared with USA I water) ), And inclusion complex of α-cyclodextrin and niacinamide (data series B; 1: 1 complex, dissolved in 0.1% w / v in medium prepared with USA I water) Mortality can be reduced. FIG. 17 shows the number of daily dead animals for each media condition and the control data series is media made with USA I water and lacks additional additives or supplements. The results show that the complexed form of α-cyclodextrin can be used as an active component of cellular hydration to delay the death of living animals.

FIG. 18 instead shows the data in FIG. 17 in a normal medium using USA I water (control) or a medium supplemented with a 1: 1 inclusion complex of α-cyclodextrin and L-arginine (sample 1). Or as survival curves for animals growing in medium supplemented with 1: 1 inclusion complex of cyclodextrin and niacinamide (Sample 2). Thus, the delay in mortality shown in FIG. 17 resulted in survival to older age, and the mean age of survival (50% survival) from approximately 13 days in normal media, cyclodextrin inclusion complex Increased to approximately 14 days in medium containing the active component of cellular hydration, indicating an 8% increase in lifespan.

While the invention has been shown and described with reference to the foregoing operational principles and preferred embodiments, various changes in form and detail will depart from the spirit and scope of the invention. It will be clear that it can be done without. The present invention is intended to embrace all such alternatives, modifications and variances that fall within the scope of the appended claims.

Claims (37)

A composition that interacts with a biological cell system comprising a biologically active molecule together with a biological molecule surface, cellular components and water molecules having a specific density,
A composition comprising a biologically active component constructed to increase the activity of a biological cell line system by increasing the hydration of one or more components of the cell line. 2. The composition of claim 1, wherein the biologically active component comprises a first carbohydrate clathrate subcomponent that increases the hydrogen bonding structure of water. 3. The composition of claim 2, wherein the biologically active component comprises a second solute subcomponent. 4. The composition of claim 3, wherein the second solute subcomponent is a bioactive agent. 2. The composition of claim 1, wherein the biologically active component comprises an inclusion complex composed of a clathrate component and a complex-forming compound. 6. The composition of claim 5, wherein the complexing compound is a bioactive agent. The composition according to claim 2, wherein the clathrate subcomponent comprises a compound selected from the group consisting of amylose and cyclodextrin. The composition of claim 2 wherein the clathrate subcomponent comprises cyclodextrin. 9. The composition of claim 8, wherein the cyclodextrin is constructed to exhibit cosmotrophic activity that increases the water binding structure. The cyclodextrin is constructed to cause an increase in hydrogen bonding between water molecules, the increase altering the hydration of the biomolecule surface of the biological cell system, thereby the cellular components of the biological cell system 10. A composition according to claim 9, which alters the interaction between. The cyclodextrin is constructed to enhance hydrogen bonding between water molecules, resulting in an open water structure having a lower specific density and increasing the diffusion rate of the bioactive molecule. The composition as described. 12. The bioactive molecule of claim 11, further comprising the bioactive molecule selected from the group consisting of enzymes, enzyme substrates, nutrients, metabolites, cytokines, neurotransmitters, hormones, extracellular signals, intracellular messengers, and pharmacological agents. Composition. The cyclodextrin is α-cyclodextrin, β-cyclodextrin and γ-cyclodextrin, methylated α-cyclodextrin, β-cyclodextrin and γ-cyclodextrin, 2-hydroxypropylated β-cyclodextrin, water-soluble Β-cyclodextrin polymer, partially acetylated α-cyclodextrin, β-cyclodextrin and γ-cyclodextrin, ethylated α-cyclodextrin, β-cyclodextrin and β-cyclodextrin, carboxyalkyl The selected β-cyclodextrin, α-cyclodextrin, β-cyclodextrin and quaternary ammonium salts of γ-cyclodextrin, amylose (e.g., acetylated amylose), and mixtures thereof. Composition. 8. The composition of claim 7, further comprising a complexing agent, wherein the clathrate subcomponent is capable of forming an inclusion complex with the complexing agent. 15. The composition of claim 14, wherein the complexing agent is selected from the group consisting of amino acids, vitamins, flavoring agents, odorants, and coloring agents. The clathrate minor component is α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, 2-hydroxypropyl-cyclodextrin, carboxymethylated cyclodextrin, quaternary ammonium cyclodextrin, amylose, amylose derivatives, and their 16. A composition according to claim 15, selected from the group consisting of a mixture. A composition that interacts with a biological cell system comprising a biologically active molecule together with a biological molecule surface, cellular components and water molecules having a specific density,
A first carbohydrate clathrate component capable of increasing the hydrogen bonding structure of water;
A second small solute component;
Including
A composition wherein the clathrate component and the solute component can increase the activity of a biological cell line by increasing the hydration of one or more components of the cell line. 18. A composition according to claim 17, wherein the clathrate subcomponent comprises a compound selected from the group consisting of amylose and cyclodextrin. 18. The composition of claim 17, wherein the second solute subcomponent is a bioactive agent. 18. The composition of claim 17, further comprising a complex-forming compound, wherein the clathrate compound and the complex-forming compound form an inclusion complex. 21. The bioactive molecule of claim 20, further comprising the bioactive molecule selected from the group consisting of enzymes, enzyme substrates, nutrients, metabolites, cytokines, neurotransmitters, hormones, extracellular signals, intracellular messengers, and pharmacological agents. Composition. The clathrate component is α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, methylated β-cyclodextrin, 2-hydroxypropylated β-cyclodextrin, water-soluble β-cyclodextrin polymer, partially acetylated Α-cyclodextrin, β-cyclodextrin and γ-cyclodextrin, ethylated α-cyclodextrin, β-cyclodextrin and γ-cyclodextrin, carboxyalkylated β-cyclodextrin, α-cyclodextrin, 19. The composition of claim 18, which is a cyclodextrin selected from the group consisting of β-cyclodextrin and γ-cyclodextrin quaternary ammonium salts, amylose (eg, acetylated amylose), and mixtures thereof. 23. The composition of claim 22, wherein the complexing agent is selected from the group consisting of amino acids, vitamins, flavoring agents, odorants, and coloring agents. The clathrate minor component is α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, 2-hydroxypropyl-cyclodextrin, carboxymethylated cyclodextrin, quaternary ammonium cyclodextrin, amylose, amylose derivatives, and their 19. A composition according to claim 18, selected from the group consisting of a mixture. A beverage that interacts with a biological cell system comprising a biologically active molecule along with a biological molecule surface, cellular components and water molecules having a specific density,
A water-containing component selected from the group consisting of a non-carbonated liquid and a carbonated liquid;
A first carbohydrate clathrate component capable of increasing the hydrogen bonding structure of water;
A second solute subcomponent,
A beverage in which the clathrate component and the solute component can increase the activity of the biological cell line by increasing the hydration of one or more components of the cell line. 26. A beverage according to claim 25, wherein the clathrate subcomponent comprises a compound selected from the group consisting of amylose and cyclodextrin. 27. A beverage according to claim 26, wherein the second solute minor component is a bioactive agent. 28. The beverage according to claim 27, further comprising a complex-forming compound, wherein the clathrate compound and the complex-forming compound form an inclusion complex. Further comprising a flavoring selected from the group consisting of apples, apricots, bananas, grapes, black currants, raspberries, peaches, pears, pineapples, peaches, oranges, and vanilla flavors, and examples of flavoring related compounds include: Butyl acetate, butyl isovalerate, allyl butyrate, amyl valerate, ethyl acetate, ethyl valerate, amyl acetate, maltol, isoamyl acetate, ethyl maltol, isomaltol, diacetyl, ethyl propionate, methyl anthranilate, methyl butyrate, butyric acid Pentyl, pentyl pentanoate, erythritol, maltitol, mannitol, sorbitol, lactitol, xylitol, inositol, isomalt, propylene glycol, glycerol (glycerin), threitol, galactitol, palatinose, Original isomalto-oligosaccharides, reduced xylo-oligosaccharides, reduced gentio- oligosaccharides, reduced maltose syrup, and reduced glucose syrup, beverage according to claim 28. Betaline, Betacyanin, Betaxanthin, Buluxanthin, Miraxanthin, Portulaxanthin, Indicaxanthin, Anthocyanidin, Aurantinidin, Cyanidin, Delphinidin, Europinidine, Luteoridinin, Pelargonidin, Malvidin, Peonidin, Petunidin, Rossinidin, Corresponding Anthocyanin or Anthocyanin 29. The beverage of claim 28, further comprising a colorant selected from the group consisting of glucoside, turmeric colorants, phenolic curcuminoids, curcumin, demethoxycurcumin and bisdemethoxycurcumin. Nicotinamide (vitamin B ₃ ), niacinamide, niacin, pyridoxal hydrochloride (vitamin B ₆ ), ascorbic acid, edible ascorbic acid ester, riboflavin, pyridoxine, thiamine, vitamin B ₉ , folic acid, folate, pteroyl-L-glutamic acid, 30. The beverage of claim 28, further comprising a vitamin selected from the group consisting of pteroyl-L-glutamate, salts thereof, and mixtures thereof. 30. A beverage according to claim 28, further comprising an amino acid or an amine. Amino acids are aspartic acid, arginine, glycine, glutamic acid, proline, threonine, theanine, cysteine, cystine, alanine, valine, tyrosine, leucine, isoleucine, asparagine, serine, lysine, histidine, ornithine, methionine, carnitine, aminobutyric acid (alpha -Isomers, beta-isomers, and gamma-isomers), glutamine, hydroxyproline, taurine, norvaline, sarcosine, salts thereof, mixtures thereof, and N-alkylated C ₁ -C of these amino acids ₃ and N- acylated C ₁ -C ₃ derivatives, and selected from the group consisting of any amino acid or a mixture of derivatives, beverage according to claim 32. A method of improving cell hydration in animals, such as humans, where physiological conditions arise and where there is a biological cell system that contains biologically active molecules along with biomolecule surfaces, cellular components and water molecules of a specific density. And
Preparing a beverage by dissolving a carbohydrate clathrate component and an inclusion complex formed by a complexing agent capable of dissociating from the carbohydrate clathrate component under physiological conditions;
Orally ingesting the beverage into an animal, wherein the carbohydrate clathrate component is dissociated from the complexing agent, altering the strength, extent, and kinetics of hydrogen-bonded water structures on the surface of the cell's biomolecules. Including the steps of: 35. The method of claim 34, further comprising selecting the clathrate component from the group consisting of amylose and cyclodextrin. The step of selecting comprises cyclodextrin as α-cyclodextrin, β-cyclodextrin, γ-cyclodextrin, methylated β-cyclodextrin, 2-hydroxypropylated β-cyclodextrin, water-soluble β-cyclodextrin polymer, Partially acetylated α-cyclodextrin, β-cyclodextrin and γ-cyclodextrin, ethylated α-cyclodextrin, β-cyclodextrin and β-cyclodextrin, carboxyalkylated β-cyclodextrin, α 36. The method of claim 35 comprising selecting from the group consisting of -cyclodextrin, β-cyclodextrin and quaternary ammonium salts of γ-cyclodextrin, amylose (eg, acetylated amylose), and mixtures thereof. 35. The method of claim 34, further comprising selecting the complexing agent from the group consisting of amino acids, vitamins, flavoring agents, odorants, and coloring agents.