METHOD AND COMPOSITIONS FOR ADMINISTERING RESVERATROL AND PTEROSTILBENE

Abstract

Resveratrol and/or pterostilbene are added to a chewable carrier, to enhance absorption of resveratrol and/or pterostilbene buccally or through the mucous membranes of the mouth. The composition containing the resveratrol and/or pterostilbene is designed to be retained in the mouth for at least 20 seconds and up to 20 minutes to ensure absorption of the resveratrol and/or pterostilbene through the mucous membranes of the mouth.

Description

CROSS-REFERENCE TO RELATED APPLICATIONS

The present application claims priority from provisional application 61/019,745, filed Jan. 8, 2008.

FIELD OF THE INVENTION

The present invention relates to methods and compositions for administering resveratrol and pterostilbene.

BACKGROUND OF THE INVENTION

Resveratrol and its analogs, pterostilbene (3,5-dimethoxy-4′-hydroxy-trans-stilbene), TMS (3,4′,5-reimwrhoxzy-trans-stilbene), 3,4′,4-DH-5-MS (3,4′-dihydroxy5-methoxy-trans-stilbnene) and 3,5-DH-4′MS(3,5-dihydroxy-4′-,ethoxy-trans-stilbene), have been shown to have chemopreventaive activity against cardiovascular disease and a variety of cancers in model systems. However, it is not clear whether the drugs reach the proposed sites of action in vivo after oral administration, especially in humans.

Walle et al., in Drug Metabolism and Disposition 32: 1377-1382, 2004, examined the absorption, bioavailability and metabolism of 14C-resveratrol after oral and intravenous doses in six human volunteers. When healthy men and women took an oral dose of 2 grams of trans-resveratrol, only traces of the unchanged resveratrol were detected in plasma. However, the resveratrol is rapidly transformed in the liver to the glucurunide, which makes the resveratrol not bioavailable.

Resveratrol is found in limited quantities in wine and some foods. The amount of resveratrol in wines ranges from about 0.05 mg/liter to about 7.13 mg/L, with red wines having the most resveratrol. Other sources of resveratrol are peanuts and peanut butter (about 1 mg per cup (about 150 g), and red grapes, which

Resveratrol and pterostilbene have been shown to have chemopreventive activity against cardiovascular disease and a variety of cancers in model systems. However, it is not clear if the drug reaches the proposed sites of action in vivo after oral ingestion, especially in humans. Unfortunately, as reported by Walle et al., Drug Metabolism and Disposition 32:1377-1382, 2004, absorption of a dietary relevant 25-mg oral dose was at least 70% with peak plasma levels of resveratrol and metabolites of 491±90 ng/ml (about 2 microM) and a plasma half-life of 9.2±0.6 hours. However, only trace amounts of unchanged resveratrol (<5 ng/ml) could be detected in plasma. Most of the oral dose was recovered in urine, and liquid chromatography/mass spectrometry analysis identified three metabolic pathways: sulfate and glucuronic acid conjugation of the phenolic groups and hydrogenation of the aliphatic double bond, the latter likely produced by intestinal microflora. Extremely rapid sulfate conjugation by the intestine/liver appears to be the rate-limiting step in the bioavailability of resveratrol and related compounds such as pterostilbene.

In order to avoid this entero-hepatic first exposure, an exposure that deactivates resveratrol, it was discovered that absorption through the buccal mucosa could bypass the enter-hepatic circulation and thus enable the resveratrol to be absorbed unchanged. Indeed, non-conjugated resveratrol levels were substantially increased in the serum in this way. It was also found that ethanol and DMSO in concentrations from 5% to about 50% could be used as vehicles or as transfer agents for resveratrol, which can increase absorption up to ten-fold.

SUMMARY OF THE INVENTION

Accordingly, Resveratrol, pterostilbene, and related compounds can be administered transmucosally or buccally by incorporating one or more of these compounds into any foodstuff that remains in the mouth for a period of time of at least five seconds or more, and preferably up to twenty minutes, such as chewing gum, taffy, hard candy, jerky, etc. Detailed Description of the Invention

Significant reductions in cardiovascular disease risk have been associated with moderate consumption of alcoholic beverages (43). This “French Paradox”, the observation that mortality from coronary heart disease is relatively low in France despite relatively high levels of dietary saturated fat and cigarette smoking, suggested that the regular consumption of red wine could provide additional protection from cardiovascular disease (44, 45). Red wine contains resveratrol and even higher levels of flavonoids. These polyphenolic compounds have antioxidant, anti-inflammatory, and other potentially anti-atherogenic effects in the test tube and in some animal modes of atherosclerosis (46). Results of epidemiological studies addressing the value of these polyphenolic compound has been inconsistent. While some large prospective studies found that wine drinkers were at lower risk of cardiovascular disease than been or liquor drinkers (47-49), others fond no difference (50-52). Several studies have discovered that people who drink wine have higher incomes, more education, smoke less, and eat more fruits and vegetables and less saturated fat than people who prefer beer or liquor (53-55). This may explain some of the differences.

In vitro, resveratrol effectively scavenges (neutralizes) free radicals an other oxidants (11) and inhibits oxidation of low density lipoprotein (12, 13). However, there is little evidence that resveratrol is an important antioxidant in vivo (14) because resveratrol is very quickly conjugated in the gastrointestinal/liver system, and intracellular concentrations of resveratrol in humans are likely to be much lower than that of other important antioxidants, such as vitamin C, vitamin E and glutathione. It is believed that the antioxidant activity of resveratrol metabolites, which comprise most of the circulating resveratrol, may be lower than that of resveratrol.

The chemical structure of resveratrol is very similar to that of the synthetic estrogen agonist, diethylstilbestrol, suggesting that resveratrol might also function as an estrogen agonist. However, in cell culture experiments resveratrol acts as an estrogen agonist under some condition, and an estrogen antagonist under other conditions (16, 17). In estrogen receptor-positive breast cancer cells, resveratrol acted as an estrogen agonist in the absence of the endogenous estrogen 17-beta-estradiol, but acted as an estrogen antagonist in the presence of 17-beta estradiol (18, 19). At present it appears that resveratrol has the potential to act as an estrogen agonist or antagonist, depending upon factors such as cell type, estrogen receptor isoform (ER alpha or ER beta), and the presence of endogenous estrogens (15).

Resveratrol, 3,4′-dihydroxystilbene, also known as 3,4′5-stilbenetriol, is a phytoalexin produced naturally by several plants when under attack by bacteria or fungi. Resveratrol has also been produced synthetically [see Farina et al., Nat. Prod. Res. 20(3): 247-252 2006]. A number of beneficial health effects of resveratrol have been reported, including anti-cancer, antiviral, neuroprotective, anti-aging, anti-inflammatory, and life-prolonging effects, although most of the studies used animal subjects rather than humans.

Resveratrol is found in widely varying amounts in grapes (primarily the skins of red grapes), raspberries, mulberries, plums, peanuts, berries of the Vaccinium species, including blueberries, bilberries, cranberries, some pines such as Scotch pine and eastern white pine, and the roots and stalks of giant knotweed and Japanese knotweed. Resveratrol was first isolated from an extract of the Peruvian legume Cassia quinquangulata in 1974.

In grapes, resveratrol is found primarily in the skin and seeds. The amount found in grape skins also varies with the grape cultivar, its geographic origin, and exposure to fungal infection. The amount of fermentation time a wine spends in contact with grape skins is an important determinant of its resveratrol content.

Pterostilbene is a stilbenoid compound chemically related to resveratrol. Other names for pterostilbene are 4-[(E)-2-(3,5-dimethoxyphenyl)ethenyl]phenol; 3′,5′-dimethoxy-4-stilbenol; and 3,5-dimethoxy-4′-hydroxy-trans-stilbene.

Pterostilbene has been found to be equal in potency to resveratrol as a cancer-preventive agent (Rimando et al., Cancer Chemopreventive and Antioxidant Activities of Pterostilbene, a Naturally Occurring Analogue of Resveratrol, Ars.usda.gov). Antioxidants destroy free radicals, highly reactive molecules whose excess has been linked to cancer, which may account for the effectiveness of resveratrol and pterostilbene against cancer. Animal studies have also found that pterostilbene can lower blood glucose and may be a potent antidiabetic agent.

Quantitative studies have shown that for every ten parts resveratrol, there are only one to two parts pterostilbene. The relationship between the two compounds and their unequal content in plants is unclear, but remains the subject of ongoing studies. Dark-skinned grapes are likely to contain the most pterostilbene. For reasons that are not clear, pterostilbene is normally not found in wine. This may be because it is unstable in light and air, which makes it less likely to survive the wine-making process.

For administration, the resveratrol and/or pterostilbene can be natural or synthetic. They may also be in the form of a pharmacologically acceptable salt, ester, amide, prodrug or analog or as a combination thereof. Salts, esters, amides, prodrugs and analogs of resveratrol and/or pterostilbene may be prepared using standard procedures known to those skilled in the art of synthetic organic chemistry and pharmaceutical formulation.

The resveratrol and/or pterostilbene can be incorporated into a chewable or other type of food or delivery vehicle that is retained in the mouth for about five seconds to about twenty minutes. Types of foods that require long chewing or that remain in the mouth for a period of time of up to twenty minutes include chewing gum, taffy, hard candies that are sucked, gummy candies, fudge, lozenges, and the like. Alternatively, the resveratrol and/or pterostilbene can be incorporated in a thin film that can be retained in the mouth and from which the active ingredient is delivered across the oral mucous membranes.

Some compounds are not carcinogenic until they have been metabolized in the body by cytochrome P450 enzymes (2). By inhibiting the expression and activity of certain cytochrome P450 enzymes (20,21), resveratrol could help prevent cancer by decreasing exposure to these activated carcinogens. In contrast, increasing the activity of phase II biotransformation enzymes generally promotes the excretion of potentially toxic or carcinogenic chemicals. Resveratrol has been found to increase the expression and activity of the phase II enzyme NAD(P)H:quinine reductase in cultured cells (5, 22).

Following DNA damage, the cell cycle can be transiently arrested to allow for DNA repair or activation of pathways leading the apoptosis if the damage is irreparable (23). Defective cell cycle regulation may result in the propagation of mutations that contribute to the development of cancer. Resveratrol has been found to induce cell cycle arrest when added to cancer cells grown in culture (24).

Unlike normal cells, cancer cells proliferate rapidly and lose the ability to respond to cell death signals which initiate apoptosis. Resveratrol has been found to inhibit proliferation and induce apoptosis in a number of cancer cell lines (21).

Cancerous cells invade normal tissue with the aid of matrix metalloproteinases. Resveratrol has been found to inhibit the activity of at least one type of matrix metalloproteinase (25). Invasive tumors must also develop new blood vessels to fuel their rapid growth by angiogenesis. Resveratrol has been found to inhibit angiogenesis in vitro (26, 27).

Inflammation promotes cellular proliferation and angiogenesis and inhibits apoptosis (28). Resveratrol has been found to inhibit the activity of several inflammatory enzymes in vitro, including cyclooxygenase and lipoxygenase (29, 30).

Atherosclerosis is now recognized as an inflammatory disease, and several measures of inflammation are associated with increased risk of myocardial infarction (31). One of the earliest events in the development of atherosclerosis is the recruitment of inflammatory while blood cells from the blood to the artery wall by vascular cell adhesion molecules (32). Resveratrol has been found to inhibit the expression of adhesion molecules in cultured endothelial cells (33, 34).

eNOS catalyzes the formation of nitric oxide (NO) by vascular endothelial cells. NO is required to maintain vasodilation, and impaired NO-dependent vasodilation is associated with increased risk of cardiovascular disease (38). Resveratrol has been found to stimulate eNOS activity in cultured endothelial cells (39, 40).

Platelet aggregation is one of the first steps in forming a blood clot that can occlude a coronary or cerebral artery, eventually resulting in myocardial infarction or stroke. Resveratrol has been found to inhibit platelet aggregation in vitro (41, 42).

Resveratrol has been found to exert a number of potentially cardioprotective effects in vitro, as discussed above, including inhibition of platelet aggregation (41, 42, 56), promotion of vasodilation by enhancing the production of NO (40, 57) and inhibiting inflammatory enzymes (30, 58, 59). However, the concentrations of resveratrol required to produce these effects are higher than those that have been measured in human plasma after oral consumption of resveratrol (7). Although the presence of resveratrol in red wine has stimulated a great deal of interest in the potential for resveratrol to prevent cardiovascular diseases, there is currently no convincing evidence that resveratrol has a cardioprotective effecting humans, particularly in the amounts present in one or two glasses of red wine daily.

When added to cells in vitro, resveratrol has been found to inhibit the proliferation of a variety of human cancer cell lines, including those from breast, prostate, stomach, colon, pancreatic and thyroid cancers (2). In animal models, oral administration of resveratrol inhibited the development of esophageal (63), intestinal (64) and mammary cancer (18, 55) induced by chemical carcinogens. It is not known whether high intakes of resveratrol can help prevent cancer in humans. Studies of human metabolism of orally ingested resveratrol may not result in tissue levels that are high enough to realize most of the protective effects demonstrate in vitro studies (7, 10). It is likely that the tissue levels of resveratrol result from the conjugation of resveratrol shortly after ingestion, which produces conjugated resveratrol that is not useful for any of the known activities of resveratrol.

As described by Rimando et al., Agric. Food Chem., 2002, 50(12):3453-3457, pterostilbene was evaluated for antioxidant potential. The peroxyl-radical scavenging activity of pterostilbene was the same as that of resveratrol, having total reactive antioxidant potentials of 237±58 and 253±53 microM, respectively. Pterostilbene exhibited significant inhibition of carcinogen-induced preneoplastic lesions using a mouse mammary organ culture model.

Accordingly, resveratrol and/or pterostilbene can be formulated in an alcoholic beverage that is retained in the mouth prior to swallowing such that the resveratrol and/or pterostilbene is absorbed through the mucous membranes of the mouth, thus bypassing the gastrointestinal/liver systems and passing directly into the blood stream. Alternatively, the resveratrol and/or ptersostilbene can be incorporated into a vehicle such as food, candy, chewing gum, films, or the like, that can be retained in the mouth for a sufficient time to deliver the resveratrol and/or ptersostilbene through the oral mucous membranes.

Rimando et al., in Agric. Food Chem., 2005, 53(9):3403-3407, reported that pterostilbene acts as a PPARA agonist and may be a more effective PPARA agonist and hypolipidemic agent than resveratrol. In vivo studies have demonstrated that pterostilbene possesses lipid and glucose lowering effects.

Hougee et al., in Planta Med, 2005, 781(5): 387-392, note that an extract of Pterocarpus marsupium Roxbn (PM) containing pterostilbene was evaluated for its PGE-2 inhibitory activity in LPS-stimulated PBMC in healthy human volunteers. The biological activity and safety of the PM extract were evaluated. It was found that PM extract, pterostilbene and resveratrol inhibited PGE2 production from LPS-stimulated human peripheral blood mononuclear cells.

Pan et al., Agric. Food Chem. 2007 September 19:55(19):7777-7785. demonstrated that pterostilbene was able to inhibit cell proliferation and induce apopotosis in a concentration- and time-dependent manner, which may provide an improved treatment of human gastric cancer.

As resveratrol and pterostilbene are rapidly conjugated when administered orally, it is desirable to administer these compounds transmucosally or buccally to avoid conjugation in the gastric-hepatic system.

Resveratrol and/or pterostilbene can be administered transmucosally or buccally by adding the resveratrol and/or pterostilbene to chewing gums or candies, or other edible substances that remain in contact with the mucous membranes of the mouth for a period of time of from about five seconds to about two minutes. The concentration should be 1-5 mg per candy or the equivalent to chewing gum or chocolate. Among the types of candies are hard candies that remain in the mouth for a prolonged period of time, and chewy candies such as taffy, gummi candies, confectionary starch and caramels, that require a long period of chewing.

One method of incorporating resveratrol and/or pterostilbene in a chewable product is to provide a consumable center and a compressible composition including the resveratrol and/or pterostilbene that is compressed around the consumable center. The resveratrol or pterostilbene may be encapsulated. The chewable center may be, by way of example and not of limitation, a gummi candy, confectionary starch, hard candy, licorice-type candy, or a tableted excipient such as dextrose, sucrose, or other saccharides, sorbitol, mannitol, iso-maltol, other sugar alcohols, or combinations thereof. The product may be in any shape and size suitable for chewing, such as a disk shape, a pellet shape, a spherical shape, or a tube shape.

Another method of incorporating resveratrol and/or pterostilbene into a chewable form for transmucosal delivery is by incorporating the resveratrol and/or pterostilbene into a dissolvable matrix material. This dissolvable matrix material may include carbohydrates, fats, proteins such as gelatin, waxes (natural and synthetic), hydrocarbons and other materials that safely dissolve in the oral cavity. The combination of resveratrol and/or pterostilbene and the matrix material can be compressed, poured into a mold cavity, dehydrated, freeze dried or otherwise formed into an integral delivery system. Specific confectionery components can be combined in order for the mixture to form an integral solid mass. These components include, for example, compressible confectioner's sugar, sorbitol, mannitol and maltodextrin. Any food-safe flavoring and/or coloring materials may be incorporated into the product.

In the formulations of resveratrol and/or pterostilbene the dissolution of the matrix material can be controlled by adding a hydrophobic agent such as calcium stearate to slow dissolution. Dissolution can also be slowed by mechanically compressing the mixture to a greater extent.

Another method for administering the resveratrol and/or pterostilbene orally by maintaining the resveratrol and/or pterostilbene in contact with the mucous membranes of the mouth is by incorporating these active ingredients in an orally nondissolvable matrix. The resveratrol and/or pterostilbene may be incorporated into a sponge-like matrix, the resveratrol and/or pterostilbene may be microencapsulated and held within a microsponge, the resveratrol and/or pterostilbene may be contained within a permeable membrane or screen-like barrier, or the resveratrol and/or pterostilbene may be held within other nondissolvable containment vehicles capable of releasing the resveratrol and/or pterostilbene for transmucosal administration.

When the resveratrol and/or pterostilbene is incorporated into a sponge-like matrix, the matrix may be designed to release the resveratrol and/or pterostilbene in response to pressure, either negative or positive, or other similar release trigger. The matrix may be held within a screen or permeable membrane which allows the resveratrol and/or pterostilbene to permeate the screen when exposed to conditions of the mouth, pharynx, or esophagus. Suitable screen-like materials include woven nylon, polypropylene or polyethylene mesh, with varying apertures or pore sizes, and porous sheet materials. A preferred screen or membrane material is preferably flexible with no or low absorption of resveratrol and/or pterostilbene and that is free of interaction with the oral mucous membranes, palatable in taste and texture, non-irritating, nontoxic, hypoallergenic and does not leach out plasticizers.

Alternatively, a sponge-like matrix may be held together with a suitable biocompatible adhesive, either dissolvable or nondissolvable. Typical adhesives include sodium carboxymethylcellulose, sodium alginate, and tragacanth.

Compositions containing resveratrol and/or pterostilbene as an active ingredient include all compositions wherein the active ingredient is contained in an amount effective to achieve its intended purpose. While individual needs vary, determination of optimal ranges of effective amounts of each compound is within the skill of the art. Typical dosages comprise 0.01 to 100 mg/kg body weight. The preferred dosages comprising 0.1 to 100 mg/kg body weight. The most preferred dosages comprise 1 to 50 mg/kg body weight.

Another vehicle for administering resveratrol and/or pterostilbene is by incorporating the resveratrol and/or pterostilbene in a thin film that dissolves slowly in the oral cavity so that the resveratrol or pterostilbene is slowly released through the mucous membranes of the mouth. These thin film products are generally formed by combining a properly selected polymer and polar solvent, as well as resveratrol and/or pterostilbene and any desired fillers.

The film-forming polymer may be water soluble, water insoluble, or a combination of one or more either water soluble or water insoluble polymers. The polymer may include cellulose or a cellulose derivative. Specific examples of useful water soluble polymers include, but are not limited to, pullulan, hydroxypropylmethylcelluilose, hydroxyethylcellulose, hydroxypropylcellulose, polyvinyl pyrrollidone, carboxymethyl cellulose, polyvinyl alcohol, sodium alginate, polyethylene glycol, xanthan gum, gum Arabic, gum tragacanth, guar gum, gum acacia, polyacrylic acid, methyl methacrylate copolymers, carboxyvinyl polymers, starch, and combinations thereof. Specific examples of useful water insoluble polymers include, but are not limited to, ethyl cellulose, hydroxypropyl ethyl cellulose, cellulose acetate phthalate, hydroxypropyl methyl cellulose phthalate and combinations thereof.

Other polymers that can be used in films for administering resveratrol and/or pterostilbene include biodegradable polymers, copolymers, block polymers and combinations thereof. Suitable polymers are described in detail in Yang et al., US Published Application No. 2003/0107149, the entire contents of which are incorporated herewith by reference. The films are preferably designed for controlled release of the resveratrol and/or pterostilbene.

Other carriers for pterostilbene and/or resveratrol are chews impregnated with an emulsion of the active ingredients. Examples of such chews can be found in Brown et al., U.S. Pat. No. 7,147,888, the entire contents of which are hereby incorporated by reference.

Pharmaceutical compositions for administering the active ingredients transmucosally or buccally as described above preferably contain, in addition to the pharmacologically active compound, suitable pharmaceutically acceptable carriers comprising excipients and auxiliaries which facilitate processing of the active compounds into preparations which can be used transmucosally or buccally.

Preferably, the dosage forms, contain from about 0.01 to about 99 percent by weight, preferably from about 20 to 75 percent by weight, active compound(s), together with the excipients. For purposes of the present invention, all percentages are by weight unless otherwise indicated. In addition to the following described pharmaceutical composition, the resveratrol and/or pterostilbene can be formulated as inclusion complexes, such as cyclodextrin inclusion complexes.

In determining the dosages of the resveratrol and/or pterostillbene to be administered, the dosage and frequency of administration is selected in relation to the pharmacological properties of the specific active ingredients. Normally, at least three dosage levels should be used. In toxicity studies in general, the highest dose should reach a toxic level but be sub lethal for most animals in the group. If possible, the lowest dose should induce a biologically demonstrable effect. These studies should be performed in parallel for each compound selected.

Additionally, the ID₅₀ level of the active ingredient in question can be one of the dosage levels selected, and the other two selected to reach a toxic level. The lowest dose that dose not exhibit a biologically demonstrable effect. The toxicology tests should be repeated using appropriate new doses calculated on the basis of the results obtained. Young, healthy mice or rats belonging to a well-defined strain are the first choice of species, an the first studies generally use the preferred route of administration. Control groups given a placebo or which are untreated are included in the tests. Tests for general toxicity, as outlined above, should normally be repeated in another non-rodent species, e.g., a rabbit or dog. Studies may also be repeated using alternate routes of administration.

Singe dose toxicity tests should be conducted in such a way that signs of acute toxicity are revealed and the mode of death determined. The dosage to be administered is calculated on the basis of the results obtained in the above-mentioned toxicity tests. It may be desired not to continue studying all of the initially selected compounds. Data on single dose toxicity, e.g., ID₅₀, the dosage at which half of the experimental animals die, is to be expressed in units of weight or volume per kg of body weight and should generally be furnished for at least two species with different modes of administration. In addition to the ID₅₀ value in rodents, it is desirable to determine the highest tolerated dose and/or lowest lethal dose for other species, i.e., dog and rabbit.

The amount of resveratrol and/or pterostilbene to be administered to any given patient must be determined empirically, and will differ depending upon the condition of the patients. Relatively small amounts of the active ingredient can be administered at first, with steadily increasing dosages if no adverse effects are noted. Of course, the maximum safe toxicity dosage as determined in routine animal toxicity tests should never be exceeded.

The foregoing description of the specific embodiments will so fully reveal the general nature of the invention that others can, by applying current knowledge, readily modify and/or adapt for various applications such specific embodiments without undue experimentation and without departing from the generic concept, and, therefore, such adaptations and modifications should and are intended to be comprehended within the meaning and range of equivalents of eth disclosed embodiments.

It is to be understood that the phraseology or terminology employed herein is for the purpose of description and not of limitation. The means and materials for carrying out various disclosed functions may take a variety of alternative forms without departing from the invention.

Thus, the expressions “means to . . . ” and “means for . . . ” as may be found in the specification above and/or in the claims below, followed by a functional statement, are intended to define and cover whatever structural, physical, chemical, or electrical element or structures which may now or in the future exist for carrying out the recited function, whether or nor precisely equivalent to the embodiment or embodiments disclosed in the specification above. It is intended that such expressions be given their broadest interpretation.

REFERENCES

• Aggarwal B B, Bhardwaj A, Aggarwal R S, Seeram N P, Shishodia S, Takada Y. Role of resveratrol in prevention and therapy of cancer: preclinical and clinical studies. Anticancer Res. 2004;24(5A):2783-2840. (PubMed)
• American Academy of Pediatrics. Committee on Substance Abuse and Committee on Children With Disabilities. Fetal alcohol syndrome and alcohol-related neurodevelopmental disorders. Pediatrics. 2000;106(2 Pt 1):358-361. (PubMed)
• Banerjee S, Bueso-Ramos C, Aggarwal B B. Suppression of 7,12-dimethylbenz(a)anthracene-induced mammary carcinogenesis in rats by resveratrol: role of nuclear factor-kappaB, cyclooxygenase 2, and matrix metalloprotease 9. Cancer Res. 2002;62(17):4945-4954. (PubMed)
• Barefoot J C, Gronbaek M, Feaganes J R, McPherson R S, Williams R B, Siegler I C. Alcoholic beverage preference, diet, and health habits in the UNC Alumni Heart Study. Am J Clin Nutr. 2002;76(2):466-472. (PubMed)
• Berge G, Ovrebo S, Eilertsen E, Haugen A, Mollerup S. Analysis of resveratrol as a lung cancer chemopreventive agent in A/J mice exposed to benzo[a]pyrene. Br J Cancer. 2004;91(7):1380-1383. (PubMed)
• Bertelli A A, Giovannini L, Giannessi D, et al. Antiplatelet activity of synthetic and natural resveratrol in red wine. Int J Tissue React. 1995;17(1):1-3. (PubMed)
• Bhat K P, Lantvit D, Christov K, Mehta R G, Moon R C, Pezzuto J M. Estrogenic and antiestrogenic properties of resveratrol in mammary tumor models. Cancer Res. 2001;61(20):7456-7463. (PubMed)
• Blake G J, Ridker P M. C-reactive protein and other inflammatory risk markers in acute coronary syndromes. J Am Coll Cardiol. 2003;41(4 Suppl S):37S-42S. (PubMed)
• Bowers J L, Tyulmenkov V V, Jernigan S C, Klinge C M. Resveratrol acts as a mixed agonist/antagonist for estrogen receptors alpha and beta. Endocrinology. 2000;141(10):3657-3667. (PubMed)
• Bradamante S, Barenghi L, Villa A. Cardiovascular protective effects of resveratrol. Cardiovasc Drug Rev. 2004;22(3):169-188. (PubMed)
• Brito P, Almeida L M, Dinis T C. The interaction of resveratrol with ferrylmyoglobin and peroxynitrite; protection against LDL oxidation. Free Radic Res. 2002;36(6):621-631. (PubMed)
• Burns J, Gardner P T, Matthews D, Duthie G G, Lean M E, Crozier A. Extraction of phenolics and changes in antioxidant activity of red wines during vinification. J Agric Food Chem. 2001;49(12):5797-5808. (PubMed)
• Burns J, Yokota T, Ashihara H, Lean M E, Crozier A. Plant foods and herbal sources of resveratrol. J Agric Food Chem. 2002;50(11):3337-3340. (PubMed)
• Carluccio M A, Siculella L, Ancora M A, et al. Olive oil and red wine antioxidant polyphenols inhibit endothelial activation: antiatherogenic properties of Mediterranean diet phytochemicals. Arterioscler Thromb Vasc Biol. 2003;23(4):622-629. (PubMed)
• Chen C K, Pace-Asciak C R. Vasorelaxing activity of resveratrol and quercetin in isolated rat aorta. Gen Pharmacol. 1996;27(2):363-366. (PubMed)
• Chen Z H, Hurh Y J, Na H K, et al. Resveratrol inhibits TCDD-induced expression of CYP1A1 and CYP1B1 and catechol estrogen-mediated oxidative DNA damage in cultured human mammary epithelial cells. Carcinogenesis. 2004;25(10):2005-2013. (PubMed)
• Ciolino H P, Yeh G C. Inhibition of aryl hydrocarbon-induced cytochrome P-450 1A1 enzyme activity and CYP1A1 expression by resveratrol. Mol Pharmacol. 1999;56(4):760-767. (PubMed)
• Creasey L L, Coffee M. Phytoalexin production potential of grape berries. J Am Soc Hortic Sci. 1988;113(2):230-234.
• Criqui M H, Ringel B L. Does diet or alcohol explain the French paradox? Lancet. 1994;344(8939-8940):1719-1723. (PubMed)
• Crowell J A, Korytko P J, Morrissey R L, Booth T D, Levine B S. Resveratrol-associated renal toxicity. Toxicol Sci. 2004;82(2):614-619. (PubMed)
• Donnelly L E, Newton R, Kennedy G E, et al. Anti-inflammatory effects of resveratrol in lung epithelial cells: molecular mechanisms. Am J Physiol Lung Cell Mol Physiol. 2004;287(4):L774-783. (PubMed)
• Duffy S J, Vita J A. Effects of phenolics on vascular endothelial function. Curr Opin Lipidol. 2003;14(1):21-27. (PubMed)
• Faxon D P, Fuster V, Libby P, et al. Atherosclerotic Vascular Disease Conference: Writing Group III: pathophysiology. Circulation. 2004;109(21):2617-2625. (PubMed)
• Ferrero M E, Bertelli A E, Fulgenzi A, et al. Activity in vitro of resveratrol on granulocyte and monocyte adhesion to endothelium. Am J Clin Nutr. 1998;68(6):1208-1214. (PubMed)
• Frankel E N, Waterhouse A L, Kinsella J E. Inhibition of human LDL oxidation by resveratrol. Lancet. 1993;341(8852):1103-1104. (PubMed)
• Fremont L. Biological effects of resveratrol. Life Sci. 2000;66(8):663-673. (PubMed)
• Fukao H, Ijiri Y, Miura M, et al. Effect of trans-resveratrol on the thrombogenicity and atherogenicity in apolipoprotein E-deficient and low-density lipoprotein receptor-deficient mice. Blood Coagul Fibrinolysis. 2004;15(6):441-446. (PubMed)
• Gehm B D, McAndrews J M, Chien P Y, Jameson J L. Resveratrol, a polyphenolic compound found in grapes and wine, is an agonist for the estrogen receptor. Proc Natl Acad Sci USA. 1997;94(25):14138-14143. (PubMed)
• German J B, Walzem R L. The health benefits of wine. Annu Rev Nutr. 2000;20:561-593. (PubMed)
• Gescher A J, Steward W P. Relationship between mechanisms, bioavailibility, and preclinical chemopreventive efficacy of resveratrol: a conundrum. Cancer Epidemiol Biomarkers Prev. 2003;12(10):953-957. (PubMed)
• Goldberg D M, Karumanchiri A, Ng E, Yan J, Eleftherios P, Soleas G. Direct gas chromatographic-mass spectrometric method to assay cis-resveratrol in wines: preliminary survey of its concentration in commercial wines. J Agric Food Chem. 1995;43(5):1245-1250.
• Goldberg D M, Yan J, Soleas G J. Absorption of three wine-related polyphenols in three different matrices by healthy subjects. Clin Biochem. 2003;36(1):79-87. (PubMed)
• Gronbaek M, Becker U, Johansen D, et al. Type of alcohol consumed and mortality from all causes, coronary heart disease, and cancer. Ann Intern Med. 2000;133(6):411-419. (PubMed)
• Haider U G, Sorescu D, Griendling K K, Vollmar A M, Dirsch V M. Resveratrol increases serine15-phosphorylated but transcriptionally impaired p53 and induces a reversible DNA replication block in serum-activated vascular smooth muscle cells. Mol Pharmacol. 2003;63(4):925-932. (PubMed)
• Hecht S S, Kenney P M, Wang M, et al. Evaluation of butylated hydroxyanisole, myo-inositol, curcumin, esculetin, resveratrol and lycopene as inhibitors of benzo[a]pyrene plus 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone-induced lung tumorigenesis in A/J mice. Cancer Lett. 1999;137(2):123-130. (PubMed)
• Heilbronn L K, Ravussin E. Calorie restriction and aging: review of the literature and implications for studies in humans. Am J Clin Nutr. 2003;78(3):361-369. (PubMed)
• Hendler S S, Rorvik D R, eds. PDR for Nutritional Supplements. Montvale: Medical Economics Company, Inc; 2001.
• Howitz K T, Bitterman K J, Cohen H Y, et al. Small molecule activators of sirtuins extend Saccharomyces cerevisiae lifespan. Nature. 2003;425(6954):191-196. (PubMed)
• Igura K, Ohta T, Kuroda Y, Kaji K. Resveratrol and quercetin inhibit angiogenesis in vitro. Cancer Lett. 2001;171(1):11-16. (PubMed)
• Jang M, Cai L, Udeani G O, et al. Cancer chemopreventive activity of resveratrol, a natural product derived from grapes. Science. 1997;275(5297):218-220. (PubMed)
• Joe A K, Liu H, Suzui M, Vural M E, Xiao D, Weinstein I B. Resveratrol induces growth inhibition, S-phase arrest, apoptosis, and changes in biomarker expression in several human cancer cell lines. Clin Cancer Res. 2002;8(3):893-903. (PubMed)
• Juan M E, Vinardell M P, Planas J M. The daily oral administration of high doses of trans-resveratrol to rats for 28 days is not harmful. J Nutr. 2002;132(2):257-260. (PubMed)
• Kirk R I, Deitch J A, Wu J M, Lerea K M. Resveratrol decreases early signaling events in washed platelets but has little effect on platelets in whole blood. Blood Cells Mol Dis. 2000;26(2):144-150. (PubMed)
• Klatsky A L, Friedman G D, Armstrong M A, Kipp H. Wine, liquor, beer, and mortality. Am J Epidemiol. 2003;158(6):585-595. (PubMed)
• Klatsky A L. Drink to your health? Sci Am. 2003;288(2):74-81. (PubMed)
• Klinge C M, Blankenship K A, Risinger K E, et al.

Resveratrol and estradiol rapidly activate MAPK signaling through estrogen receptors alpha and beta in endothelial cells. J Biol Chem. 2004. (PubMed)

• Li Z G, Hong T, Shimada Y, et al. Suppression of N-nitrosomethylbenzylamine (NMBA)-induced esophageal tumorigenesis in F344 rats by resveratrol. Carcinogenesis. 2002;23(9):1531-1536. (PubMed)
• Lin M T, Yen M L, Lin C Y, Kuo M L. Inhibition of vascular endothelial growth factor-induced angiogenesis by resveratrol through interruption of Src-dependent vascular endothelial cadherin tyrosine phosphorylation. Mol Pharmacol. 2003;64(5):1029-1036. (PubMed)
• Lin S J, Defossez P A, Guarente L. Requirement of NAD and SIR2 for life-span extension by calorie restriction in Saccharomyces cerevisiae. Science. 2000;289(5487):2126-2128. (PubMed)
• Lu R, Serrero G. Resveratrol, a natural product derived from grape, exhibits antiestrogenic activity and inhibits the growth of human breast cancer cells. J Cell Physiol. 1999;179(3):297-304. (PubMed)
• McCann S E, Sempos C, Freudenheim J L, et al. Alcoholic beverage preference and characteristics of drinkers and nondrinkers in western New York (United States). Nutr Metab Cardiovasc Dis. 2003;13(1):2-11. (PubMed)
• Meng X, Maliakal P, Lu H, Lee M J, Yang C S. Urinary and plasma levels of resveratrol and quercetin in humans, mice, and rats after ingestion of pure compounds and grape juice. J Agric Food Chem. 2004;52(4):935-942. (PubMed)
• Mnjoyan Z H, Fujise K. Profound negative regulatory effects by resveratrol on vascular smooth muscle cells: a role of p53-p21(WAF1/CIP1) pathway. Biochem Biophys Res Commun. 2003;311(2):546-552. (PubMed)
• Moreno-Labanda J F, Mallavia R, Perez-Fons L, Lizama V, Saura D, Micol V. Determination of piceid and resveratrol in Spanish wines deriving from Monastrell (Vitis vinifera L.) grape variety. J Agric Food Chem. 2004;52(17):5396-5403. (PubMed)
• Mortensen E L, Jensen H H, Sanders S A, Reinisch J M. Better psychological functioning and higher social status may largely explain the apparent health benefits of wine: a study of wine and beer drinking in young Danish adults. Arch Intern Med. 2001;161(15):1844-1848. (PubMed)
• Mukamal K J, Conigrave K M, Mittleman M A, et al. Roles of drinking pattern and type of alcohol consumed in coronary heart disease in men. N Engl J Med. 2003;348(2):109-118. (PubMed)
• Pace-Asciak C R, Hahn S, Diamandis E P, Soleas G, Goldberg D M. The red wine phenolics trans-resveratrol and quercetin block human platelet aggregation and eicosanoid synthesis: implications for protection against coronary heart disease. Clin Chim Acta. 1995;235(2):207-219. (PubMed)
• Pinto M C, Garcia-Barrado J A, Macias P. Resveratrol is a potent inhibitor of the dioxygenase activity of lipoxygenase. J Agric Food Chem. 1999;47(12):4842-4846. (PubMed)
• Piver B, Berthou F, Dreano Y, Lucas D. Inhibition of CYP3A, CYP1A and CYP2E1 activities by resveratrol and other non volatile red wine components. Toxicol Lett. 2001;125(1-3):83-91. (PubMed)
• Regev-Shoshani G, Shoseyov O, Kerem Z. Influence of lipophilicity on the interactions of hydroxy stilbenes with cytochrome P450 3A4. Biochem Biophys Res Commun. 2004;323(2):668-673. (PubMed)
• Renaud S C, Gueguen R, Siest G, Salamon R. Wine, beer, and mortality in middle-aged men from eastern France. Arch Intern Med. 1999;159(16):1865-1870. (PubMed)
• Rimando A M, Kalt W, Magee J B, Dewey J, Ballington J R. Resveratrol, pterostilbene, and piceatannol in vaccinium berries. J Agric Food Chem. 2004;52(15):4713-4719. (PubMed)
• Rimm B E, Klatsky A, Grobbee D, Stampfer M J. Review of moderate alcohol consumption and reduced risk of coronary heart disease: is the effect due to beer, wine, or spirits. BMJ. 1996;312(7033):731-736. (PubMed)
• Romero-Perez A I, Ibern-Gomez M, Lamuela-Raventos R M, de La Torre-Boronat M C. Piceid, the major resveratrol derivative in grape juices. J Agric Food Chem. 1999;47(4):1533-1536. (PubMed)
• Romero-Perez A I, Lamuela-Raventos R M, Waterhouse A L, de la Torre-Boronat M C. Levels of cis- and trans-resveratrol and their glucosides in white and rose Vitis vinifera wines from Spain. J Agric Food Chem. 1996;44(8):2124-2128.
• Sanders T H, McMichael R W, Jr., Hendrix K W. Occurrence of resveratrol in edible peanuts. J Agric Food Chem. 2000;48(4):1243-1246. (PubMed)
• Schneider Y, Duranton B, Gosse F, Schleiffer R, Seiler N, Raul F. Resveratrol inhibits intestinal tumorigenesis and modulates host-defense-related gene expression in an animal model of human familial adenomatous polyposis. Nutr Cancer. 2001;39(1):102-107. (PubMed)
• Siemann E H, Creasey L L. Concentration of the phytoalexin resveratrol in wine. Am J Enol Vitic. 1992;43(1):49-52.
• Sobolev V S, Cole R J. trans-resveratrol content in commercial peanuts and peanut products. J Agric Food Chem. 1999;47(4):1435-1439. (PubMed)
• Soleas G J, Diamandis E P, Goldberg D M. Resveratrol: a molecule whose time has come? And gone? Clin Biochem. 1997;30(2):91-113. (PubMed)
• St Leger A S, Cochrane A L, Moore F. Factors associated with cardiac mortality in developed countries with particular reference to the consumption of wine. Lancet. 1979;1(8124):1017-1020. (PubMed)
• Steele V E, Hawk E T, Viner J L, Lubet R A. Mechanisms and applications of non-steroidal anti-inflammatory drugs in the chemoprevention of cancer. Mutat Res. 2003;523-524:137-144. (PubMed)
• Stewart Z A, Westfall M D, Pietenpol J A. Cell-cycle dysregulation and anticancer therapy. Trends Pharmacol Sci. 2003;24(3):139-145. (PubMed)
• Stocker R, Keaney J F, Jr. Role of oxidative modifications in atherosclerosis. Physiol Rev. 2004;84(4):1381-1478. (PubMed)
• Stojanovic S, Sprinz H, Brede O. Efficiency and mechanism of the antioxidant action of trans-resveratrol and its analogues in the radical liposome oxidation. Arch Biochem Biophys. 2001;391(1):79-89. (PubMed)
• Szewczuk L M, Forti L, Stivala L A, Penning T M. Resveratrol is a peroxidase-mediated inactivator of COX-1 but not COX-2: a mechanistic approach to the design of COX-1 selective agents. J Biol Chem. 2004;279(21):22727-22737. (PubMed)
• Tangkeangsirisin W, Serrero G. Resveratrol in the chemoprevention and chemotherapy of breast cancer. In: Bagchi D, Preuss H G, eds. Phytopharmaceuticals in Cancer Chemoprevention. Boca Raton: CRC Press; 2005:449-463.
• Tessitore L, Davit A, Sarotto I, Caderni G. Resveratrol depresses the growth of colorectal aberrant crypt foci by affecting bax and p21(CIP) expression. Carcinogenesis. 2000;21(8):1619-1622. (PubMed)
• Tsai S H, Lin-Shiau S Y, Lin J K. Suppression of nitric oxide synthase and the down-regulation of the activation of NFkappaB in macrophages by resveratrol. Br J Pharmacol. 1999;126(3):673-680. (PubMed)
• Walle T, Hsieh F, Delegge M H, Oatis J E, Jr., Walle U K. High absorption but very low bioavailability of oral resveratrol in humans. Drug Metab Dispos. 2004;32(12):1377-1382. (PubMed)
• Wallerath T, Deckert G, Ternes T, et al. Resveratrol, a polyphenolic phytoalexin present in red wine, enhances expression and activity of endothelial nitric oxide synthase. Circulation. 2002;106(13):1652-1658. (PubMed)
• Wang Z, Huang Y, Zou J, Cao K, Xu Y, Wu J M. Effects of red wine and wine polyphenol resveratrol on platelet aggregation in vivo and in vitro. Int J Mol Med. 2002;9(1):77-79. (PubMed)
• Wang Z, Zou J, Huang Y, Cao K, Xu Y, Wu J M. Effect of resveratrol on platelet aggregation in vivo and in vitro. Chin Med J (Engl). 2002;115(3):378-380. (PubMed)
• Wannamethee S G, Shaper A G. Type of alcoholic drink and risk of major coronary heart disease events and all-cause mortality. Am J Public Health. 1999;89(5):685-690. (PubMed)
• Wilson T, Knight T J, Beitz D C, Lewis D S, Engen R L. Resveratrol promotes atherosclerosis in hypercholesterolemic rabbits. Life Sci. 1996;59(1):PL15-21. (PubMed)
• Woo J H, Lim J H, Kim Y H, et al. Resveratrol inhibits phorbol myristate acetate-induced matrix metalloproteinase-9 expression by inhibiting JNK and PKC delta signal transduction. Oncogene. 2004;23(10):1845-1853. (PubMed)
• Wood J G, Rogina B, Lavu S, et al. Sirtuin activators mimic caloric restriction and delay ageing in metazoans. Nature. 2004;430(7000):686-689. (PubMed)
• Yang S H, Kim J S, Oh T J, et al. Genome-scale analysis of resveratrol-induced gene expression profile in human ovarian cancer cells using a cDNA microarray. Int J Oncol. 2003;22(4):741-750. (PubMed)
• Ziegler C C, Rainwater L, Whelan J, McEntee M F. Dietary resveratrol does not affect intestinal tumorigenesis in Apc(Min/+) mice. J Nutr. 2004;134(1):5-10. (PubMed)

Claims

1. A composition for administering at least one compound selected from the group consisting of resveratrol and, pterostilbene comprising adding the at least one compound to a carrier that requires chewing or retention in the mouth for from about 20 seconds to about twenty minutes.

2. The composition according to claim 1 wherein the carrier is selected from the group consisting of chewing gum, gummi candy, taffy, caramel candy, fudge, degradable thin films, nondegradable thin films and hard candy.

3. The composition according to claim 1 wherein the at least one compound is present in an amount to provide a dose of from about 1 g to about 5 g per piece of carrier.

4. A method for administering at least one compound selected from the group consisting of resveratrol and pterostilbene, their esters or any such conjugate that does not eliminate their bio activity, that are absorbable via the mouth mucosa membranes to an individual in need thereof comprising:

a. incorporating the at least one compound into a carrier that requires chewing or retention in the mouth;

b. administering the carrier to the individual so that the individual retains the carrier in the mouth;

c. whereby the at least one compound is absorbed through the mucous membranes of the mouth.

5. The method according to claim 4 wherein the carrier is retained in the mouth over a period of from about 20 seconds to about 20 minutes.

6. The method according to claim 4 wherein the carrier is selected from the group consisting of chewing gum, gummy candy, taffy, caramel, candy, fudge, degradable thin films, nondegradable thin films, and hard candy.