WO2017217868A1

WO2017217868A1 - Water-dispersible propolis composition

Info

Publication number: WO2017217868A1
Application number: PCT/NZ2016/050124
Authority: WO
Inventors: Andrew George Lavrent
Original assignee: Individual
Current assignee: Individual
Priority date: 2016-06-16
Filing date: 2016-08-04
Publication date: 2017-12-21
Anticipated expiration: 2018-12-16

Abstract

The invention relates to a water-dispersible propolis composition, methods of making the same, and aqueous formulations comprising the composition, including food, beverage, nutraceutical, pharmaceutical and cosmetic formulations.

Description

WATER-DISPERSIBLE PROPOLIS COMPOSITION

1. FIELD OF THE INVENTION

The invention relates to a propolis composition that is water-dispersible. 2. BACKGROUND OF THE INVENTION

Propolis (Honey bee glue) is the generic name for a product made by honey bees (Apis mellifera). It is obtained from resinous exudates of plants, bark and leaf buds, which are collected and processed by bees by the addition of salivary secretions and wax. This material is used to coat parts of the interior of the hive, sealing any cracks present. Bees collect resins from numerous plant species. The chemical composition of propolis varies greatly. It depends directly on the local flora and phenology of the host plants and indirectly on the locality and time of collection. It is believed that some of the substances collected undergo chemical alteration by bee enzymes prior to incorporation into propolis, making propolis a substance of both plant and animal origin. Specifically, bees mix the plant exudates (including resins, waxes and oils collected from branches, flowers, pollen and buds) with beeswax secreted through glands located near the hypopharygeal region of the bees' bodies. The addition of β-glucosidase secreted during plant exudate collection and small amounts of sugar, results in whole propolis.

Crude propolis typically consists of resin (20-55%), wax (30-40%), volatile oils (5-10%), and various phenolic compounds (10-30%), including, in particular, flavonoids, aromatic carboxylic acids and aromatic aldehydes. The phenolic portion also comprises phenylalkyl acids, such as cinnamic acid, caffeic acid, feruiic acid and p-coumaric acid and some esters thereof.

Over 150 compounds have been identified in a single propolis sample with more than 180 compounds isolated from propolis samples to date. These compounds are believed responsible for propolis's numerous biological and pharmacological properties, including antibacterial, antiviral, antifungal, anti-inflammatory and antioxidant properties.

Propolis-based preparations have been known for a long time in popular medicine and nutraceuticals. Their main applications relate to treatment of respiratory tract infections, flu, acne, wounds, burns, herpes, gingivitis and stomatitis, as well as prevention of caries.

Propolis-based preparations have also been used in the cosmetics industry for the formulation of creams, serums, ointments, shampoos, lotions and gels.

Propolis has a unique aromatic smell and differs in colour depending on the plant sources. However, crude propolis is a blackish, brown mass consisting mostly of hydrophobic substances. It smells and tastes unpleasant. Therefore, it is typically processed before use.

The main processing step applied to crude propolis is the removal of the wax component, which has no biological activity and makes the propolis difficult to handle.

Solvent extraction of the propolis with a water-soluble organic solvent, either before or after dewaxing, provides a usable propolis formulation. Propolis formulations for use in the nutraceutical or cosmetics industry are generally in the form of alcoholic extracts.

At room temperature, propolis extracts usually appear as brown/red to dark green/brown substances with a sticky and ductile consistency. Their complicated chemical composition and sticky, non-wettable properties make them difficult to use. For example, propolis components tend to fall out of solution when currently available alcoholic or propylene glycol propolis preparations are added to an aqueous medium. A resinous substance of insoluble components forms around the edge of the container. This greatly limits the applications that propolis can be used in.

Ethoxylated surfactants such as alcohol ethoxylates and alcohol ethoxysulfates, for example, sodium laureth sulfate, can be added to increase the dispersibility of an alcoholic propolis extract in aqueous solution. However, these agents are undesirable in most propolis applications such as their use as food or beverage ingredients, or as agents in nutraceutical, pharmaceutical, or cosmetic formulations.

A more water-dispersible composition could be obtained by extraction with a more polar solvent, for example, 20% ethanol in water. However, the increase in water-dispersibility arises only because the proportion of lipophilic components in the extract has decreased . As the efficacy of propolis is thought to be mostly due to the lipophilic components, this method does not provide a biologically active propolis product.

Therefore, there is a need for a propolis composition that retains the efficacy of crude propolis but is water-dispersible. It is an object of the invention to provide such a composition, or to at least provide the public with a useful choice.

3. SUMMARY OF THE INVENTION

In one aspect the invention provides a water-dispersible propolis composition comprising (a) about 0.01 to about 75 wt% propolis substance and

(b) about 25 to about 99.09 wt% dispersant, wherein the dispersant comprises cocoyi proline and at least one polyglycerol ester.

In another aspect the invention provides a method for making a water-dispersible propolis composition comprising mixing about 0.01 to about 75 wt% propolis substance with about 25 to about 99.09 wt% dispersant under heated conditions, wherein the dispersant comprises cocoyi proline and at least one polyglycerol ester, and wherein the relative amounts of propolis substance and dispersant do not exceed 100 wt%.

In one embodiment, the propolis substance and dispersant are mixed at 30 to about 105°C, preferably 65 to 80°C.

In one embodiment the propolis substance and dispersant are mixed under vacuum.

In one aspect, the invention provides an aqueous formulation comprising a water- dispersible propolis composition of the invention.

In one embodiment, the aqueous formulation is a food or beverage formulation. In another embodiment, the aqueous formulation is a nutraceutical or pharmaceutical formulation.

In another embodiment, the aqueous formulation is a cosmetic formulation. In the above aspects:

In one embodiment the propolis substance is a water-miscible organic solvent extract of crude propolis. Preferably, the crude propolis is de-waxed .

In one embodiment the water-miscible organic solvent extract of crude propolis contains less than about 2 wt% solvent.

In one embodiment, the water-miscible organic solvent is ethanol.

In another embodiment, the ratio of cocoyi proline to at least one polyglycerol ester in the dispersant is about 2: 1 to about 1 :4, preferably about 1 : 1 to about 1 : 3, more preferably about 1 : 2.

In another embodiment the at least one polyglycerol ester comprises polyglycerol-6 caprylate. 4. DETAILED DESCRIPTION OF THE INVENTION

4.1 Definitions

As used herein, the term "dispersion stability" refers to the ability of a substance to remain dispersed when present in an aqueous solution or formulation. A substance with high dispersion stability will remain dispersed for several months. A substance with low dispersion stability will precipitate, aggregate or otherwise fall out of solution shortly or immediately after being mixed with an aqueous formulation.

As used herein, the term "under heated conditions" means that the components are heated either before, during or after mixing. As used herein, the term "comprising" means "consisting at least in part of". When interpreting statements in this specification and claims which include the term

"comprising", other features besides the features prefaced by this term in each statement can also be present. Related terms such as "comprise" and "comprised" are to be interpreted in similar manner. 4.2 The composition of the invention

In one aspect the invention provides a water-dispersible propolis composition comprising

(a) about 0.01 to about 75 wt% propolis substance and

(b) about 25 to about 99.09 wt% dispersant, wherein the dispersant comprises cocoyl proline and at least one polyglycerol ester. The propolis substance for use in the invention comprises crude propolis or any extract thereof. The propolis utilized in the invention can be from any geographical location including but not limited to New Zealand, Brazil, China, Japan, Africa, Australia, Europe and United States of America .

The term "crude propolis" as used herein refers to raw propolis from which the plant and bee detritus such as sticks, wings and other debris has been removed.

In one embodiment the crude propolis is de-waxed. De-waxed crude propolis can be produced by heating raw propolis and skimming off the detritus and melted wax.

A propolis extract can be prepared by mixing de-waxed crude propolis with a solvent in which some of the propolis components dissolve, and then collecting the solvent. Water- miscible organic solvents are preferred, as they have the correct polarity to extract the beneficial phenolic compounds present in the crude propolis. When de-waxed crude propolis is mixed with a water-miscible organic solvent, an upper layer containing propolis components will separate from a lower layer containing viscous sediment. The upper layer, which can be concentrated by evaporation, distillation or other standard techniques, constitutes the propolis extract. Examples of water miscible solvents for use in the invention include but are not limited to alcohols such as methanol, ethanol, n-propanol, isopropanol, ethylene glycol, diethylene glycol, propylene glycol, dipropylene glycol and other glycols; ketone solvents such as acetone and methyl ethyl ketone; and ether solvents such as ethylene glycol and dimethyl ether. Where the propolis substance for use in the composition of the invention is a propolis extract, it should be substantially free of the solvent used to extract the propolis components. Less than 2 wt% solvent is preferred. Without being bound by theory it is thought that excess solvent may interfere with the interaction between the propolis components and the dispersant, reducing the dispersibility of the composition. In one embodiment the propolis substance is a water-miscible organic solvent extract of crude propolis. Preferably, the crude propolis is de-waxed .

In one embodiment, the water-miscible organic solvent is ethanol. In another embodiment, the water-miscible organic solvent is aqueous ethanol comprising at least about 50 wt% ethanol, preferably at least about 60 wt% ethanol, more preferably, at least about 70 wt% ethanol.

In one embodiment, the water-dispersible propolis composition comprises about 0.01 to about 60 wt% propolis substance, preferably about 0.01 to about 45 wt%, more preferably about 0.01 to about 30 wt%.

In the composition of the invention the propolis substance is mixed with a dispersant comprising cocoyl proline and at least one polyglycerol ester.

As would be understood by a person skilled in the art, the relative amounts of propolis substance and dispersant cannot exceed 100 wt%. Cocoyl proline is a lipoamino acid of vegetable origin which is obtained by acylation of coconut fatty acids with proline.

Polyglycerol esters are generally prepared by esterification of selected polyglycerol molecules with fatty acids or by alcoholysis of a vegetable oil with a polyglycerol. Their wide HLB (hydrophilic/lipophilic balance) range makes them suitable for both w/o and o/w systems. As such, they are extensively used in the food industry as solubilizers, emulsifiers, clouding agents, crystallization inhibitors, and suspending agents. Polyglycerol esters for use in the composition of the invention include but are not limited to polyglyceryl-2 caprylate, polyglyceryl-4 caprylate, polyglyceryl-3 caprylate, polyglyceryl-6 caprylate, polyglyceryl-10 caprylate, polyglyceryl-4 caprylate/ca prate, polyglyceryl-5 caprylate/caprate, polyglyceryl-6 caprylate/caprate, polyglyceryl-7 caprylate/ca prate, polyglyceryl-8 caprylate/caprate, polyglyceryl-9 caprylate/caprate, polyglyceryl-10 caprylate/caprate, polyglyceryl-4 caprate, polyglyceryl-5 caprate, polyglyceryl-6 caprate, polyglyceryl-7 caprate, polyglyceryl-8 caprate, polyglyceryl-9 caprate, polyglyceryl-10 caprate, polyglyceryl-4 laurate, polyglyceryl-5 laurate, polyglyceryl-6 laurate, polyglyceryl- 7 laurate, polyglyceryl-8 laurate, polyglyceryl-9 laurate, polyglyceryl-10 laurate, polyglyceryl-6 cocoate, polyglyceryl-7 cocoate, polyglyceryl-8 cocoate, polyglyceryl-9 cocoate, polyglyceryl-10 cocoate, polyglyceryl-11 cocoate, polyglyceryl-12 cocoate, polyglyceryl-6 myristate, polyglyceryl-7 myristate, polyglyceryl-8 myristate, polyglyceryl-9 myristate, polyglyceryl-10 myristate, polyglyceryl-11 myristate, polyglyceryl-12 myristate, polyglyceryl-10 oleate, polyglyceryl-11 oleate, polyglyceryl-12 oleate, polyglyceryl-10 stearate, polyglycerol polyricinoleate, polyglyceryl-11 stearate, polyglyceryl-12 stearate, and combinations of two or more thereof.

In one embodiment, the weight ratio of cocoyl proline to at least one polyglycerol ester in the dispersant is about 2: 1 to 1 :4, preferably about 1 : 1 to 1 : 3, more preferably about 1 : 2.

In the composition of the invention, the cocoyl proline and at least one polyglycerol ester together interact with the propolis substance to give a composition in which the propolis components remain in solution, even when the composition is added to an aqueous phase.

Accordingly, the composition of the invention provides a water-dispersible form of propolis. When combined with or introduced to an aqueous medium, it displays high dispersion stability.

This dispersion stability is only achieved when both the cocoyl proline and at least one polyglycerol ester are used together, indicating a synergistic interaction between the components. Mixing either of the components with a propolis substance separately does not provide a water-dispersible propolis composition. Nor can the effect be replicated by replacing the cocoyl proline with an alternative lipoamino acid.

In one embodiment, the propolis composition also comprises one or more additional agents selected from the group comprising antioxidants, stabilisers, taste-imparting agents, color- imparting agents, flavor-imparting agents and fillers.

Examples include but are not limited to vitamins such as tocopherol, carotenoid, L-ascorbic acid, a-glucosyl L-ascorbic acid, rutin and a-glycosyl rutin; organic acids such as citric acid and malic acid; and saccharides such as glucose, maltose, lactose and maltitol. In one embodiment, the propolis composition of the invention is free from ethoxylated solubilizers and sulfated anionic surfactants.

Examples of different water-dispersible compositions of the invention are provided in Table 1 below.

Table 1

In one aspect, the invention provides a water-dispersible propolis composition comprising (a) about 0.01 to about 75 wt% propolis substance, and (b) about 25 to about 99.09 wt% dispersant, wherein the dispersant comprises cocoyl proline and at least one polyglycerol ester in a weight ratio of about 2: 1 to 1 :4. The propolis composition of the invention can be used in combination with other substances in a variety of compositions such as foods and beverages, nutraceuticals, pharmaceuticals and cosmetics, as discussed below.

The water-dispersibility of the composition means that it can be added to aqueous formulations without the propolis components coming out of solution.

4.3 The process for making the water-dispersible propolis composition of the invention

In one aspect the invention provides a method for making a water-dispersible propolis composition comprising mixing about 0.01 to about 75 wt% propolis substance with about 25 to about 99.09 wt% dispersant under heated conditions, wherein the dispersant comprises cocoyl proline and at least one polyglycerol ester, and wherein the relative amounts of propolis substance and dispersant do not exceed 100 wt%. The components can be mixed together to form the propolis composition using any suitable technique in the art. Simple mechanical mixing or agitation is sufficient to produce the composition of the invention.

The components are generally heated while being mixed, preferably under vacuum. The mixture of components can be heated at a temperature of about 30°C to about 105°C, preferably at a temperature between about 65°C and 80°C.

Once mixed, the composition is allowed to cool to room temperature.

4.4 Aqueous formulation comprising the propolis composition of the invention

The water-dispersible propolis composition of the invention can be used as an ingredient in a variety of formulations. While it is compatible with non-aqueous formulations, it is highly advantageous in aqueous formulations, in which conventional propolis compositions are generally not stable.

In one aspect, the invention provides an aqueous formulation comprising a water- dispersible propolis composition of the invention. In one embodiment, an aqueous formulation of the invention comprises about 0.001 wt% to about 60 wt% of a water-dispersible propolis composition of the invention. Preferably, the formulation comprises about 0.01 wt% to about 40 wt% of a water-dispersible propolis composition of the invention, more preferably, about 0.01 wt% to about 15 wt%.

In one embodiment, the aqueous formulation comprises about 1 wt% to about 15 wt%.

An aqueous formulation of the invention can be prepared by mixing a water-dispersible propolis composition of the invention with an aqueous phase using a high speed mixing device until the mixture is clear. In one embodiment, the water-dispersible propolis composition of the invention is mixed with an aqueous phase at a temperature from -2°C - 95°C, preferably at a temperature of about 18°C to about 30°C, at a pH of about 5 to about 8.

The preparation of the aqueous formulation and its classification is directly affected by the equipment used. Higher speed processing provides an aqueous formulation in which the propolis particles are smaller.

For example, use of a high speed blender would achieve a polydispersion; use of high shear disruption (HSD) would likely achieve a microdispersion or colloidal dispersion, while processing via ultra high pressure (UHPH) or high pressure homogenisation (HPH) would achieve a microdispersion, colloidal dispersion or nanodispersion. The aqueous formulation of the invention could also be prepared using colloidal mill, ultrasonication (UH) or membrane disruption and the like.

The water-dispersible propolis composition of the invention has a satisfactory color, flavor and taste, so can be added to foods and beverages, either directly, or as an aqueous formulation comprising the water-dispersible composition of the invention. It harmonizes with other substances having their own salty, astringent, or bitter taste, so is useful as a flavor-stabilizing agent.

Its water-solubility makes it ideal for beverages such as fruit drinks and teas.

It can also be used to improve the shelf-life and/or quality of specific health-foods, as well as a variety of general food products, for example, seasonings, Japanese-style

confectioneries, Western-style confectioneries, sherbets, ice creams, beverages, spreads, pastes, pickled products, bottled and canned products, processed meat and fish meat products, processed marine products, processed milk and egg products, processed vegetable products, processed fruit products and processed cereal products. The water-dispersible propolis composition of the invention can be incorporated into pet foods and feeds for animals, such as domestic animals, poultry, bees, silkworms and fish.

Accordingly, in one aspect, the invention provides a food or beverage formulation comprising a water-dispersible propolis composition of the invention.

The water-dispersible propolis composition of the invention can also be used in a cosmetic formulation. Cosmetic formulations of the invention include, but are not limited to, toothpastes, toothgels, lipsticks, lip-creams, sun-screening agents, concealers,

foundations, mascaras, shampoos, lotions, bath salts, refining agents, skin-whitening agents and hair restorers.

A toothpaste formulation of the invention could include about 0.8-1.2 wt% water- dispersible propolis composition mixed with excipients such as sorbitol, glycerin, calcium carbonate, lauryl glycoside, carrageenan, xanthan gum, peppermint oil, teat tree oil, menthol, spearmint oil, polyglycerol-10 laurate, liquorice, sodium benzoate and xylitol.

A toothgel formulation could include 0.8-1.2 wt% water-dispersible propolis composition mixed with excipients such as sorbitol, xylitol, glycerin, hydrophilic formed silica, carrageenan, xanthan gum and sodium benzoate.

Accordingly, in one aspect, the invention provides a cosmetic formulation comprising a water-dispersible propolis compound of the invention. Aqueous formulations of the invention may include but are not limited to lotions, creams, gels, sticks, sprays, shaving creams, ointments, cleansing liquid washes and solid bars, shampoos, pastes, powders, mousses, shaving creams, wipes, patches, nail lacquers, wound dressing, adhesive bandages, hydrogels, films. These products may comprise several types of

pharmaceuticaiiy-acceptabie carriers including, but not limited to solutions, emulsions (e.g., microemulsions and nanoemulsions), gels, solids, micelles, and liposomes.

In one aspect, the invention provides a nutraceutical or pharmaceutical composition comprising a water-dispersible propolis compound of the invention .

In one embodiment, the nutraceutical or pharmaceutical composition may be in the form of a solid, paste and liquid, for example, a troche, sublingual tablet, parenteral formulation, cachou, gargle or any other convenient form.

A cosmetic, nutraceutical or pharmaceutical formulation of the invention may be

formulated for topical application to skin. In one embodiment, a topical composition of the invention includes an aqueous formulation comprising a water-dispersible propolis composition of the invention, and a pharmaceutically acceptable topical carrier. In one embodiment, the pharmaceuticaiiy-acceptabie topical carrier comprises from about 50% to about 99.99%, by weight, of the composition (e.g., from about 80% to about 95%, by weight, of the composition.

The topical compositions useful in the present invention can be formulated as solutions. Solutions typically include an aqueous solvent (e.g., from about 50% to about 99.99%, such as from about 90% to about 99%, by weight of a pharmaceutically acceptable aqueous solvent).

Topical compositions of the invention may be formulated as a solution comprising an emollient. Such compositions preferably contain from about 2% to about 50% of an emollient(s). As used herein, "emollients" refer to materials used for the prevention or relief of dryness, as well as for the protection of the skin. A wide variety of suitable emollients are known and may be used herein. See the International Cosmetic Ingredient Dictionary and Handbook, eds. Wenninger and McEwen, pp. 1656-61, 1626, and 1654-55 (The Cosmetic, Toiletry, and Fragrance Assoc., Washington, D.C., 7^th Edition, 1997) (hereinafter "ICI Handbook") contains numerous examples of suitable materials.

A lotion can be made from such a solution. Lotions typically comprise from about 1% to about 20% (e.g., from about 5% to about 10%) of an emollient(s) and from about 50% to about 90% (e.g., from about 60% to about 80%) of water.

Another type of product that may be formulated from a solution is a cream. A cream typically comprises from about 5% to about 50% (e.g., from about 10% to about 20%) of an emollient(s) and from about 45% to about 85% (e.g., from about 50% to about 75%) of water. A recipe for a cream formulation comprising a water-dispersible propolis composition of the invention is provided in the table below Propolis Cream

De-ionised water 60.00 - 71.35 %

Caprylic/Capric Triglyceride 4.500 - 5.250 %

Helianthus Annuus (Sunflower) Seed Oil 2.500 - 3.95 %

Cetearyl alcohol and cetearyl glucoside 2.500 - 3.95 %

Water, Pentylene Glycol and Glycerin

Sodium Lactate, Lactic Acid, Serine, Uream

Sorbitol, Sodium Chloride and Allantoin 2.500 - 3.95 %

Simmondsia Chinensis (Jojoba) Seed Oil 2.500 - 3.95 %

Glycerine 2.500 3.95 %

Cetearyl wheat bran glycosides / Cetearyl alcohol 1.850 2.500 %

Water Dispersible Propolis Extract 1.250 3.000 %

Butyrospermum Parkii (Shea Butter) 1.250 3.000 %

Biosaccharide Gum 1.000 2.250 %

Arachidyl Alcohol, Behenyl Alcohol and Arachidyl Glucoside 1.000 2.250 %

Shea Butter 0.500 1.250 %

Tocopherol Acetate 0.500 0.750 %

Carbomer 0.300 0.500 %

Xanthan Gum 0.300 0.500 %

Isodecyl Citrate 0.250 0.450 %

Phenoxyethanol 0.250 0.450 %

Sodium Benzoate 0.100 0.250%

Potassium Benzoate 0.100 0.250%

Xanthan Gum 0.100 0.250%

Yet another type of product that may be formulated from a solution is an ointment. An ointment may comprise a simple base of animal or vegetable oils or semi-solid

hydrocarbons. An ointment may comprise from about 2% to about 10% of an emollient(s) plus from about 0.1% to about 2% of a thickening agent(s). A more complete disclosure of thickening agents or viscosity increasing agents useful herein can be found in the ICI Handbook pp. 1693-1697. The topical compositions useful in the present invention can also be formulated as emulsions. If the carrier is an emulsion, from about 1% to about 10% (e.g., from about 2% to about 5%) of the carrier comprises an emulsifier(s). Emulsifiers may be nonionic, anionic or cationic. Suitable emulsifiers are disclosed in the ICI Handbook, pp.1673-1686.

Lotions and creams can be formulated as emulsions. Typically, such lotions comprise from 0.5% to about 5% of an emulsifier(s). Such creams would typically comprise from about 1% to about 20% (e.g., from about 5% to about 10%) of an emollient(s); from about 20% to about 80% (e.g., from 30% to about 70%) of water; and from about 1% to about 10% (e.g., from about 2% to about 5%) of an emulsifier(s).

Single emulsion skin care preparations, such as lotions and creams, of the oil-in-water type and water-in-oil type are well-known in the cosmetic art and are useful in the subject invention. Multiphase emulsion compositions, such as the water-in-oil-in-water type are also useful in the subject invention. In general, such single or multiphase emulsions contain water, emollients, and emulsifiers as essential ingredients.

The topical compositions of this invention can also be formulated as a gel (e.g., an aqueous gel using a suitable gelling agent(s)). Suitable gelling agents for aqueous gels include, but are not limited to, natural gums, acrylic acid and acrylate polymers and copolymers, and cellulose derivatives (e.g., hydroxymethyl cellulose and hydroxypropyl cellulose). Suitable gelling agents for oils (such as mineral oil) include, but are not limited to, hydrogenated butylene/ethylene/styrene copolymer and hydrogenated

ethylene/propylene/styrene copolymer. Such gels typically comprises between about 0.1% and 5%, by weight, of such gelling agents.

The topical compositions of the present invention can also be formulated into a solid formulation (e.g., a wax-based stick, soap bar composition, powder, or a wipe containing powder).

Liposomal formulations are also useful compositions of the subject invention. Examples of liposomes are unilamellar, multilamellar, and paucilamellar liposomes, which may or may not contain phospholipids. Liposomes typically have size from about 50 nm to about 10 microns, such as about 0.1 to about 1 microns. Such compositions can be prepared by first combining hesperetin with a phospholipid, such as dipalmitoylphosphatidyl choline, cholesterol and water. Epidermal lipids of suitable composition for forming liposomes may be substituted for the phospholipid. Example of such epidermal lipids include, but are not limited to, glyceryl monoesters and diesters, polyethylene fatty ethers, and sterols. The liposome preparation may then incorporated into one of the above carriers (e.g., suspended in a solution, gel, or an oil-in-water emulsion) in order to produce the liposomal formulation. Micelle formulations are also useful compositions of the subject invention. Such

compositions can be prepared using single chain surfactants and lipids. Micelles typically have size from about 1 nm to about 100 nm, such as from about 10 nm to about 50 nm. The micelle preparation may then incorporated into one of the above carriers (e.g., a gel or a solution) in order to produce the micelle formulation.

The topical compositions useful in the subject invention may contain, in addition to the aforementioned components, a wide variety of additional oil-soluble materials and/or water-soluble materials conventionally used in compositions for use on skin, hair, and nails at their art-established levels. 5. EXAMPLES

Example 1: Preparation of a water-dispersible propolis composition of the invention

An ethanolic propolis extract (25g) was combined with 25g cocoyl proline and 50g polyglycerol-6 caprylate. The mixture was heated at about 65°C to 80°C, under mechanical agitation at medium speed.

The composition produced was then cooled to ~20°C to 25°C to give water-dispersible propolis composition A of the invention (see Table 1).

Example 2: Preparation of an aqueous formulation comprising a water- dispersible propolis composition of the invention

Water-dispersible propolis composition A, as described in Table 1 was combined with water, at concentrations of 4.0 and 12.0 wt% as set out in Table 2.

The mixtures were placed in suitable vessels, then mixed using mechanical

agitation/stirring to ensure a homogeneous mix. Using an IKA Eurostar 60 - standard impeller at 800 - 1000 RPM, the components were mixed for 8 minutes between 25°C and 30°C. Both formulations (3% and 1%) demonstrated good dispersion stability, ie they remained dispersed after 7 days.

The formulations are designated 1% and 3%, reflecting the percentage of propolis substance present in the formulation. Table 2: Aqueous formulations of the invention (800-1000 RPM)

The process was repeated, using 12.0 and 5 wt% of water-dispersible propolis composition A as set out in Table 3. Mixing was achieved using an IKA Magic lab Ultra Turrax T25 at high speed/high shear disruptor 20,000 -22,000 min-1 for 5 minutes at about 25°C to 30°C. Both formulations were stable translucent solutions.

Table 3: Aqueous formulations of the invention (20K to 22K RPM)

The process was repeated, using 12.0 and 4 wt% of water-dispersible propolis composition A, as set out in Table 4. However, these preparations were prepared using a ultra-high pressure homogenizer/microfluidizer - Avestin Emulsiflex C5 - Homogenizing pressure ~120 MPa -outlet temperature controlled by heat exchanger -30°C -35°C. Both formulations were translucent solutions with nano-solution characteristics.

Table 4: Aqueous formulations of the invention (ultra-high P)

Formulation Water-dispersible Water Zetasizer

propolis comp Measurement

(wt %)

3% 12.0 88.0 10 -100 nm

Formulation

1% 4.0 96.0 10 -100 nm

Formulation The process was repeated, suing 12.0 and 4.0 wt% of water-dispersible propolis composition A, as set out in Table 5. Mixing was achieved using mechanical agitation ensure a homogeneous mix - Ultrasonic homogenizer /sonicator. Both formulations were translucent solutions with evident nano characteristics.

Table 5:

Example 3: Stability of aqueous formulations of water-dispersible propolis compositions

A prescribed amount of a water-dispersible composition of the invention prepared in accordance with Example 2 was placed in a beaker and mixed with water to decrease the concentration of the aqueous formulation to 10, 8, 5, 3, 2, 1, and 0.50 w/w % water- dispersible propolis composition. The resultant solutions were successively allowed to stand at 50°C for 3 hours. The propolis component remained in solution in all the formulations.

Example 4: A mineral water beverage comprising a water-dispersible propolis composition of the invention

Mineral water with a mineralization (i.e., the sum of the concentrations of anions and cations present in the water) of at least about 200 mg/L (e.g., from about 300 mg/L to about 1000 mg/L) was combined with water-dispersible propolis of the invention. The mineral water comprised at least about 10 mg/L of calcium and/or at least about 5 mg/L of magnesium and included flavour compounds and sweetening agents. Generally, about 0.001 wt% to about 25 wt% of water-dispersible propolis composition was used, preferably from about 0.001 wt% to about 10 wt%.

Example 5: An energy drink comprising a water-dispersible propolis composition of the invention

Water was added to some or all of the following compounds: caffeine or guarana /gotu kola nut extract (caffeine Ξ 80 - 250 mg ), taurine glucuronolactone , citrus aurantium (synepherine ), natural sugarcane juice with polisweet (policosanols), camellia sinens, yerba mate, cordyceps, ginko biloba, ginseng, manuka honey, kiwi seed extract, grape seed extract, arginine pyroglutamic acid, n-acetyl tyrosine, lysine, choline, inositol, ethyl/calcium pyruvate, calcium phosphate /potassium phosphate water and oil soluble vitamins including flavour compounds and sweetening agents. Generally, the water- dispersible propolis is present at about 0.001 wt% to about 25 wt%, preferably from about 0.001 wt% to about 10 wt%.

It is not the intention to limit the scope of the invention to the abovementioned examples only. As would be appreciated by a skilled person in the art, many variations are possible without departing from the scope of the invention.

Claims

What we claim is:

1. A water-dispersible propolis composition comprising

(a) about 0.01 to about 75 wt% propolis substance and

2. A composition according to claim 1 which comprises about 0.01 to about 60 wt% propolis substance.

3. A composition according to claim 1 which comprises about 0.01 to about 45 wt% propolis substance.

4. A composition according to claim 1 which comprises about 0.01 to about 30 wt % propolis substance.

5. A composition according to any one of claims 1-4 wherein the ratio of cocoyi proline to at least one polyglycerol ester in the dispersant is about 2: 1 to about 1 :4.

6. A composition according to claim 5 wherein the ratio of cocoyi proline to at least one polyglycerol ester in the dispersant is about 1 : 1 to about 1 : 3.

7. A composition according to claim 6 wherein the ratio of cocoyi proline to at least one polyglycerol ester in the dispersant is about 1 : 2.

8. A composition according to any one of claims 1 to 7 wherein the at least one polyglycerol ester comprises polyglycerol-6 caprylate.

9. A composition according to any one of claims 1 to 8 wherein the propolis substance comprises a water-miscible organic solvent extract of crude propolis.

10. A composition according to claim 9 wherein the water-miscible organic solvent extract of crude propolis contains less than 2 wt% solvent.

11. A composition according to claim 9 or claim 10 wherein the water-miscible organic solvent is ethanol.

12. A composition according to any one of claims 9-11, wherein the water-miscible organic solvent is aqueous ethanol comprising about 50 wt% ethanol.

13. A composition according to any one of claims 9-11, wherein the water-miscible organic solvent is aqueous ethanol comprising about 60 wt% ethanol.

14. A composition according to any one of claims 9-11, wherein the water-miscible organic solvent is aqueous ethanol comprising about 70 wt% ethanol.

15. A composition according to any one of claims 9-14 wherein the crude propolis is de- waxed.

16. A method for making a water-dispersible propolis composition comprising mixing about 0.01 to about 75 wt% propolis substance with about 25 to about 99.09 wt% dispersant under heated conditions, wherein the dispersant comprises cocoyi proline and at least one polyglycerol ester, and wherein the relative amounts of propolis substance and dispersant do not exceed 100 wt%.

17. A method according to claim 16 wherein the propolis substance comprises a water- miscible organic solvent extract of crude propolis.

18. A method according to claim 17 wherein the water-miscible organic solvent extract of propolis contains less than 2 wt% solvent.

19. A method according to claim 17 or claim 18 wherein the water-miscible organic solvent is aqueous ethanol.

20. A method composition according to any one of claims 17-19 wherein the crude propolis is de-waxed.

21. A method according to any one of claims 16 to 20 wherein the propolis substance and dispersant are mixed at 30°C to about 105°C.

22. A method according to claim 21 wherein the propolis substance and dispersant are mixed at 65°C to 80°C.

23. A method for making a water-dispersible propolis composition according to any one of claims 16 to 22 wherein the propolis substance and dispersant are mixed under vacuum.

24. An aqueous formulation comprising a water-dispersible propolis composition of any one of claims 1 to 15.

25. An aqueous formulation according to claim 24 which comprises about 0.001 to about 60 wt% of the water dispersible propolis composition.

26. An aqueous formulation according to claim 24 or claim 25 which is a food or beverage formulation.

27. An aqueous formulation according to claim 24 or claim 25 which is a nutraceutical or pharmaceutical formulation.

28. An aqueous formulation according to claim 24 or claim 25 which is a cosmetic formulation.