Transmucosal delivery of tocotrienols

Abstract

The present invention relates to pharmaceutical compositions formulated for transmucosal delivery and in particular sublingual delivery, which pharmaceutical compositions include at least one tocotrienol or its derivatives, together with one or more pharmaceutically acceptable excipients. The present invention further relates to the use of said compositions for treating or preventing post-exercise muscle soreness, delayed muscle soreness, myocardial fibrosis, hypertension, inflammation, stroke, cancer, high cholesterol and / or triglycerides, alopecia, hypertrophy, conditions caused by radiation exposure, stabilizing and / or controlling blood sugar levels, and improving exercise endurance and performance.

Description

[0001] This application is a divisional application of an invention patent application with an application date of November 13, 2013, application number 201380022566.8, and invention name “Transmucosal delivery of tocotrienols”. Technical Field

[0002] The present invention relates to pharmaceutical compositions for transmucosal (eg, buccal, sublingual and nasal) administration comprising tocotrienols and derivatives thereof. Background Art

[0003] Vitamin E is an essential nutrient for the body. It is composed of four tocopherols (α, β, γ, δ) and four tocotrienols (α, β, γ, δ). The difference between tocotrienols and tocopherols is that for tocotrienols, the unsaturated side chain has three double bonds in its farnesyl isoprenoid tail, while in tocopherols these double bonds are single bonds ( Figure 1 ).

[0004] Tocotrienols are found in selected vegetable oils (such as palm oil and rice bran oil), certain types of fruit (such as annatto and saw palmetto), nuts (such as macadamia nuts), and plant products (such as rubber tree latex). The tocotrienol component of total vitamin E is generally lower than the tocopherol component.

[0005] Chemically, tocotrienols and each of the tocopherol isomers have antioxidant capacity due to their ability to donate a hydrogen atom (proton plus electron) from the hydroxyl group on the chromanol ring to free radicals in the body. This process deactivates ("quenches") the free radical by effectively donating a single unpaired electron (along with the hydrogen atom) to the residue.

[0006] Vitamin E has long been known for its antioxidant properties against lipid peroxidation in biological membranes and alpha-tocopherol was previously thought to be the most active form. However, in vivo, tocotrienols are stronger antioxidants, and the lipid oxygen free radical absorbance capacity (ORAC) value of delta-tocotrienol is the highest. Recent data indicate that tocotrienols are better antioxidants than tocopherols in preventing cardiovascular disease and cancer, and in treating diabetes. Current formulations of vitamin E supplements are primarily composed of alpha-tocopherol.

[0007] In addition to its fat-soluble antioxidant properties, tocotrienols have many uses. Tocotrienols specifically inhibit cholesterol biosynthesis via the liver through enhanced degradation by HMG-CoA reductase (Song et al. "Insig dependent ubiquitination and degradation of 3-hydroxy-3-methylglutarylcoenzyme a reductase by delta- and gamma-tocotrienols" The Journal of Biological Chemistry 281(35):25054-61). Tocotrienols have been shown to inhibit inflammatory pathways mediated by NF-κΒ (Nesaretnam et al. "Tocotrienols: inflammation and cancer" Ann N Y Acad Sci. 2011 July; 1229: 18-22). Tocotrienols have also been identified as agonists of peroxisome proliferator-activated receptors (PPARs), particularly PPAR-γ, which, in addition to increasing adipogenesis, is an insulin-sensitizer (Fang et al. "Vitamin E tocotrienols improve insulin sensitivity through activating peroxisome proliferator-activated receptors" Mol Nutr Food Res 2010 March; 54(3): 345-52). Indeed, tocotrienols affect many more biochemical pathways than tocopherol and are being developed as treatments for inflammation, ischemia-related diseases (e.g., stroke and myocardial infarction), dyslipidemia, and even cancer (Khosia et al. "Postprandial levels of the natural vitamin Etocotrienol in human circulation," Antioxidants & Redox Signalling 8(5-6): 1059-68).

[0008] Tocotrienols have been shown or have the potential to:

[0009] Possesses strong antioxidant properties (Serbinova et al. "Free radical recycling and intramembrane mobility in the antioxidant properties of alpha-tocopherol and aplpha-tocotrienol" Free Radical Biology & Medicine 10(5):263-75).

[0010] • Reversal of hypertension and myocardial fibrosis (Black et al. "Palm tocotrienols protect ApoE+ / - mice from diet induced atheroma formation" J Nutrition 2000; 130(10):2420-6).

[0011] Improved glycemic control and insulin response (Kuhad et al. (2009) "Suppression of NF-κβ signaling pathway by tocotrienol can prevent diabetes associated cognitive defects." Pharmacology Biochemistry, and Behaviour 92(2):251-9)

[0012] · Inhibit cholesterol biosynthesis specifically via the liver, i.e., they can lower cholesterol levels and improve dyslipidemia (Song et al. "Insig dependent ubiquitination and degradation of 3-hydroxy-3-methylglutaryl coenzyme a reductase by delta- and gamma-tocotrienols" The Journal of Biological Chemistry 281(35):25054-61)

[0013] Inhibit inflammatory pathways mediated by cyclooxygenase-2 and 12-lipoxygenase, i.e. they can be used to treat inflammation (Khanna et al. "Molecular basis of vitamin E action: tocotrienol modulates 12-lipoxygenase, a key mediator of glutamate induced neurodegeneration" J Biol Chem 2003; 278: 43508-43515)

[0014] Potentially useful for treating stroke, myocardial infarction, and even cancer (Hussein et al. "d-Delta-tocotrienol-mediated suppression of the proliferation of human PANC-1, MIA PaCa-2, and BxPC-3 pancreatic carcinoma cells" Pancreas 38(4): e124-36)

[0015] Improved exercise endurance and improved muscle glycogen levels (Lee et al. "Effects of tocotrienol-rich fraction on exercise endurance capacity and oxidative stress in forced swimming rats" Eur J Appl Physiol 2009; 107(5):587-95)

[0016] As a radioactive countermeasure for humans exposed to radiation (Ghosh et al. "Gamma-tocotrienol, atocol antioxidant as a potent radioprotector" Int J Radiat Biol 85(7):598-606)

[0017] Dietary lipids and fat-soluble vitamins must first be emulsified by bile and packaged into micelles for transport to the circulation to be absorbed from the gastrointestinal tract. Bile secretion depends on the level and type of dietary fat consumed, and studies have shown that tocotrienol absorption is reduced in fasting individuals relative to satiated individuals (Yap et al. "Pharmacokinetics and biovailability of alpha-.gamma-and delta-tocotrienols under different food status" J Pharm Pharmacol 2001 January; 53(1): 67-71). Therefore, oral administration of tocotrienols alone by tube feeding or gel capsules may lack sufficient fat content to stimulate sufficient bile secretion into the small intestine, which would be necessary to promote tocotrienol absorption.

[0018] After oral administration, tocotrienols are absorbed from the intestine and transported to the systemic circulation via the lymphatic pathway. Studies in humans have shown that the relative bioavailability of gamma-tocotrienol increases when given with food and that plasma tocotrienol concentrations do not increase significantly after tocotrienol ingestion in fasting humans (Yap et al. "Pharmacokinetics and biovailability of alpha-.gamma-and delta-tocotrienols under different food status" J Pharm Pharmacol 2001 January; 53(1): 67-71). Although food enhances gamma-tocotrienol absorption by stimulating the secretion of bile and pancreatic enzymes that enhance the formation of mixed micelles, gamma-tocotrienol absorption is still limited and far from sufficient.

[0019] In this way, it is very difficult to obtain and / or maintain the therapeutic level of gamma-tocotrienol in blood and target tissues by simple oral administration, because the absorption and transportation mechanism in the body are extremely limited and show a significant preference to alpha-tocopherol. Although the various products comprising tocotrienols are commercially available, these products are capsules that are simply filled with the mixture of various tocopherols and tocotrienol oils and are sold as the nutritional supplement for oral consumption. This type of preparation or delivery system shows poor solubility in the liquid of the intestinal tract and the high oral dose of tocotrienols suppresses its own absorption from the digestive tract. So, when simply formulated as the capsule delivery system filled with oil, only relatively low-level tocotrienols will arrive in the blood, and therefore, a strategy currently used is to use emulsifiers to enhance the absorption from the gastrointestinal tract.

[0020] Tocotrienols can be associated with lipoprotein particles called chylomicrons and absorbed via the digestive tract lacteal system, in which tocotrienols are transported to circulation via the lymphatic system. From here, the level absorbed by the tissue changes. Some reports point out that most of them end in skin and adipose tissue, wherein lower absorption enters other tissues (Gee (Ji), PT, "Unleashing the untold and misunderstood observations on vitamin E (explaining the undisclosed and misunderstood comments on vitamin E)" Genes & Nutrition (genes and nutrition) February 2011, volume 6, the 1st issue, pages 5-16). Tocotrienols can be incorporated into very low density lipoproteins and repackaged as lipoproteins at least in part by being incorporated into the alpha-tocopherol transporter mediated to be absorbed into the liver, for exporting to other tissues via circulation. For example, gamma-tocotrienols and delta-tocotrienols seem to have very low levels absorbed by key metabolic tissues (such as skeletal muscle and liver).

[0021] Tocotrienol supplementation does not seem to confer a therapeutic effect in medium-sized clinical trials in patients with dyslipidemia, regardless of the fact that plasma tocotrienols are elevated by oral supplementation. This may be due to inappropriate dosage, the competitive effect of the α-tocopherol supplemented together, or the insufficient level of tocotrienols present in the liver, so as to inhibit the biosynthesis of cholesterol. This means that reports of positive effects are limited to rodent studies and some small human trials. It has therefore been suspected whether the beneficial effects found in animal studies can be translated to humans, and whether the absorption / storage of tocotrienols is lacking. It is also clear that α-tocopherol has the highest affinity to transporters (called α-tocopherol transporter-α-TTP) and tocotrienols seem to be more rapidly metabolized than α-tocopherol, which may be due to its unsaturated isoprenoid tail and the reduced stability caused by α-TTP (due to competition from α-tocopherol). It is believed that the lack of any beneficial effects observed in these studies is due to the poor bioavailability of orally delivered tocotrienols, or to the competing effects of alpha-tocopherol present in these compositions, or a combination of these two factors.

[0022] In view of the potential clinical benefits of tocotrienols and their low toxicity (Nakamura et al., "Oral Toxicity of a tocotrienol preparation in rats," Food Chem Toxicol 2001 Aug;39(8):799-805), it would be useful to provide formulations of tocotrienols having a higher bioavailability than has heretofore been possible. Attempts have been made to improve the bioavailability of tocotrienols by incorporating them into multilamellar vesicles with lipid nanoparticles or transferrin, which appear to enhance the antitumor effects of tocotrienols by up to 70-fold (Fu et al., "Novel tocotrienol-entrapping vesicles can eradicate solid tumours after intravenous administration," J Control Release 2011 Aug 25;154(1):20-6). However, such formulations are limited because they must be introduced intravenously (which is not practical or suitable for nonclinical applications and has limited market acceptance for all but the most serious and life-threatening therapeutic indications) and rely on the use of tocopherol-based multilamellar vesicles, which themselves may interfere with the activity of the tocotrienols present.

[0023] The availability and absorption of tocotrienols into key metabolic tissues (e.g., muscle and liver) via oral administration are relatively low. In a recent paper, supplementing 400 mg of tocotrienols daily for 12 weeks only reached lower or sub-nanomoles / g levels in tissues, and the blood concentration was still below 2umol / L in all males. (Patel et al., "Oral tocotrienols are transported to human tissues and delay the progression of a model of end-stage liver disease", Journal of Nutrition 2012, 142 (3): 512-519). Therefore, there is a need for methods of delivering higher levels of tocotrienols to these tissues while minimizing metabolic degradation by the liver. Summary of the invention

[0024] The present inventors have discovered that administration of tocotrienols via a transmucosal delivery route provides enhanced bioavailability when compared to oral administration of tocotrienols.

[0025] Thus, the present invention relates to a pharmaceutical composition formulated for transmucosal delivery comprising at least one tocotrienol or a derivative thereof together with one or more pharmaceutically acceptable excipients.

[0026] In a preferred embodiment, transmucosal delivery is sublingual or buccal delivery.

[0027] In a further preferred embodiment, the composition comprises at least one tocotrienol or a derivative thereof and at least one tocopherol, wherein the tocopherol component is less than the tocotrienol component.

[0028] In a further preferred embodiment, the composition comprises at least one tocotrienol or a derivative thereof in the form of a tocotrienol-rich fraction of a vitamin E extract wherein the alpha-tocopherol component is less than the tocotrienol component.

[0029] In a further preferred embodiment, the composition includes a tocopherol and the ability of the tocopherol to interfere with or compete with the therapeutic activity of the tocotrienol or its derivative has been eliminated, reduced or minimized.

[0030] The present invention further relates to a method of treating or preventing a disease or condition amenable to treatment with a tocotrienol, the method comprising transmucosally administering at least one tocotrienol or a derivative thereof.

[0031] In a preferred embodiment, the tocotrienol or derivative thereof is administered sublingually or buccally.

[0032] In a further preferred embodiment, the tocotrienol or derivative thereof is administered in a pharmaceutical tablet, film, wafer, gum, powder, spray, solution or gel formulation.

[0033] In a further preferred embodiment, the disease or condition is selected from the group consisting of post-exercise muscle soreness, delayed onset muscle soreness, myocardial fibrosis, hypertension, inflammation, stroke, cancer, high cholesterol and / or triglycerides, hair loss and a condition caused by radiation exposure.

[0034] The present invention further relates to a method for improving exercise tolerance comprising transmucosally administering at least one tocotrienol or a derivative thereof.

[0035] The present invention further relates to a method of promoting weight and body fat loss comprising transmucosally administering at least one tocotrienol or a derivative thereof.

[0036] The present invention further relates to a method for stabilizing and / or controlling blood glucose levels in an animal, the method comprising transmucosally administering at least one tocotrienol or a derivative thereof.

[0037] The present invention further relates to a transmucosal pharmaceutical composition comprising at least one tocotrienol or a derivative thereof for use in the prevention or treatment of a disease or condition amenable to treatment with tocotrienols.

[0038] In a preferred embodiment, the transmucosal pharmaceutical composition is formulated for sublingual or buccal administration.

[0039] In a further preferred embodiment, the transmucosal pharmaceutical composition is in the form of a tablet, film, wafer, gum, powder, spray, solution or gel.

[0040] The present invention further relates to a transmucosal pharmaceutical composition comprising a tocotrienol-rich vitamin E fraction wherein the alpha-tocopherol component is less than the tocotrienol component.

[0041] In a further preferred embodiment, the transmucosal pharmaceutical composition comprises a tocotrienol-rich fraction wherein the alpha-tocopherol component constitutes no more than about 10% of the vitamin E fraction.

[0042] In a further preferred embodiment, the disease or condition is selected from the group consisting of post-exercise muscle soreness, delayed onset muscle soreness, myocardial fibrosis, hypertension, inflammation, stroke, cancer, high cholesterol and / or triglycerides, hair loss and a condition caused by radiation exposure.

[0043] The present invention further relates to a transmucosal pharmaceutical composition comprising at least one tocotrienol or a derivative thereof for use in improving exercise endurance in animals.

[0044] The present invention further relates to a transmucosal pharmaceutical composition comprising at least one tocotrienol or a derivative thereof for use in promoting weight loss in animals.

[0045] The present invention further relates to a transmucosal pharmaceutical composition comprising at least one tocotrienol or a derivative thereof for use in stabilizing and / or controlling blood glucose levels in animals.

[0046] The present invention further relates to the use of at least one tocotrienol or a derivative thereof for the preparation of a medicament for transmucosal administration.

[0047] The present invention further relates to tocotrienols for use in transmucosal administration.

[0048] The present invention further relates to the use of at least one tocotrienol or a derivative thereof in the preparation of a medicament for the prevention or treatment of a disease or condition amenable to treatment with tocotrienol, wherein the medicament is formulated for transmucosal administration.

[0049] In a preferred embodiment, the medicament is formulated for sublingual or buccal administration.

[0050] In a further preferred embodiment, the medicament is formulated for sublingual administration and is in the form of a tablet, film, wafer, gum, powder, spray, solution or gel.

[0051] The present invention further relates to a pharmaceutical composition for treating or preventing a disease or condition amenable to treatment with tocotrienols, the pharmaceutical composition comprising a therapeutically effective amount of at least one tocotrienol or a derivative thereof and one or more excipients suitable for transmucosal pharmaceutical administration.

[0052] In a preferred embodiment, the pharmaceutical composition is formulated for sublingual or buccal administration.

[0053] In a further preferred embodiment, the pharmaceutical composition is formulated for sublingual administration and is in the form of a tablet, film, wafer, gum, powder, spray, solution or gel.

[0054] The present invention further relates to a method of increasing the bioavailability of tocotrienols administered to an animal, the method comprising transmucosally administering at least one tocotrienol or a derivative thereof.

[0055] The present invention further relates to a method of minimizing the dose of tocotrienol required to achieve a therapeutic effect in an animal, the method comprising transmucosally administering at least one tocotrienol or a derivative thereof.

[0056] The present invention further relates to a transmucosal pharmaceutical composition comprising at least one tocotrienol or a derivative thereof for use in increasing the bioavailability of tocotrienol administered to an animal.

[0057] The present invention further relates to a transmucosal pharmaceutical composition comprising at least one tocotrienol or a derivative thereof for use in minimizing the dosage required to achieve a therapeutic effect by administering the tocotrienol in an animal.

[0058] The present invention further relates to the use of a transmucosal pharmaceutical composition comprising at least one tocotrienol or a derivative thereof for preventing or treating a disease or condition amenable to treatment with tocotrienols.

[0059] In a preferred embodiment, the transmucosal pharmaceutical composition is formulated for sublingual or buccal administration.

[0060] In a further preferred embodiment, the transmucosal pharmaceutical composition is in the form of a tablet, film, wafer, gum, powder, spray, solution or gel.

[0061] In a further preferred embodiment, the transmucosal pharmaceutical composition comprises a tocotrienol-rich vitamin E portion, wherein the alpha-tocopherol component constitutes up to about 10% of the vitamin E portion.

[0062] In a further preferred embodiment, the disease or condition is selected from the group consisting of post-exercise muscle soreness, delayed onset muscle soreness, myocardial fibrosis, hypertension, inflammation, stroke, cancer, high cholesterol and / or triglycerides, hair loss and a condition caused by radiation exposure.

[0063] The present invention further relates to the use of a transmucosal pharmaceutical composition for improving exercise endurance in animals, the transmucosal pharmaceutical composition comprising at least one tocotrienol or a derivative thereof.

[0064] The present invention further relates to the use of a transmucosal pharmaceutical composition for promoting weight loss in an animal, the transmucosal pharmaceutical composition comprising at least one tocotrienol or a derivative thereof.

[0065] The present invention further relates to the use of a transmucosal pharmaceutical composition comprising at least one tocotrienol or a derivative thereof for stabilizing and / or controlling blood glucose levels in an animal.

[0066] Throughout the present description and claims, the terms "comprise", "comprises", "comprising" and "comprised" are not intended to exclude additional components, integers or steps, unless the context requires otherwise.

[0067] Reference to any prior art in this specification is not, and should not be taken as, an acknowledgment or any form of suggestion that such prior art forms part of the common general knowledge in any jurisdiction or that such prior art could reasonably be expected to be recognised, understood and regarded as relevant by a person skilled in the art. BRIEF DESCRIPTION OF THE DRAWINGS

[0068] Figure 1 : The structures of the most common tocopherols and tocotrienols are shown. The main tocotrienols differ from tocopherols only in that the former have an unsaturated isoprenoid tail. In addition to the structures shown here, there are also some novel tocotrienols isolated from, for example, kiwi fruit.

[0069] Figure 2 : In Sprague-Dawley rats with obesity induced by feeding a high-fat (22%) diet for 8 weeks, dietary tocotrienols (25 mg / kg / d) improved tolerance to a glucose load (2 g / kg, ip) compared to animals not receiving tocotrienols (n=9, p<0.01; Mathai et al., 2011).

[0070] Figure 3 : In Sprague-Dawley rats maintained on a high-fat diet for 8 weeks, dietary tocotrienols (50 mg / kg / d) gradually reversed established hypertension compared with a placebo control group.

[0071] Figure 4 :exist Figure 3 The reduction in blood pressure observed in the study was accompanied by a reduction in myocardial fibrosis, as measured by left ventricular hydroxyproline content.

[0072] Figure 5 : A separate experiment showed that myocardial fibrosis induced by isoproterenol was inhibited by tocotrienol supplementation. Since this model of myocardial fibrosis did not increase blood pressure, the antifibrotic effect of tocotrienols was shown to be independent of the antihypertensive effect.

[0073] Figure 6 : Dietary supplementation with tocotrienols increases renal sodium excretion in rats fed a high-fat diet. This is important because obesity is often accompanied by sodium and water retention, which can lead to comorbidities such as elevated blood volume and blood pressure and cardiovascular disease.

[0074] Figure 7 : Effects of tocotrienols on exercise endurance in rats. A: Maximum rate of depletion of administered tocotrienols (TCT) in rats relative to controls. B: Time to depletion of administered tocotrienols (TCT) in rats relative to controls.

[0075] Figure 8 : Effects of tocotrienols on exercise endurance in humans. Time to exhaustion after cycling was measured in subjects given a sublingual tablet comprising tocotrienols relative to controls.

[0076] Fig. 9: Apparent plasma concentrations of gel and tablet sublingual formulations of tocotrienols using a 40 mg dose. Plasma levels of delta tocotrienol (delta-T3) and gamma tocotrienol (γ-T3) were measured at time points up to 180 minutes from administration of the gel or tab formulations.

[0077] Fig.10 : Delayed onset muscle soreness based on the visual analogue scale on days 1 and 2 after eccentric forearm contractions. DETAILED DESCRIPTION

[0078] The inventors have unexpectedly discovered that transmucosal delivery of tocotrienols results in enhanced bioavailability and efficacy of tocotrienols over orally delivered tocotrienols.

[0079] Transmucosal and particularly sublingual delivery of tocotrienols avoids first-pass degradation of tocotrienols by the liver, thereby increasing the absorption of tocotrienols into target tissues including skeletal muscle, adipose tissue, pancreas, heart and brain as well as liver and vascular endothelial tissue.

[0080] Many factors can affect the suitability of a particular compound for transmucosal (e.g., sublingual) delivery, including, but not limited to, the lipophilicity of the compound of interest and whether the compound has a favorable oil-water partition coefficient. An oil-water partition coefficient in the range of 40 to 2000 is considered optimal for sublingual delivery of a drug. Given that tocotrienols are highly lipophilic, delivery of tocotrienols by a transmucosal route, and particularly a sublingual route (where all membranes are coated with an aqueous saliva / mucosal layer as a barrier to contact and access), was considered an unfeasible delivery method prior to the development of the present invention.

[0081] Tocotrienols or derivatives thereof that can be used in the compositions and methods of the present invention include naturally occurring tocotrienols (extracted from natural sources) and synthetic tocotrienols. Naturally occurring tocotrienols include α, β, γ and δ tocotrienols. Although naturally occurring tocotrienols are known to exist in only one stereoisomeric form, other stereoisomers can be produced synthetically.

[0082] Derivatives of tocotrienol include, but are not limited to, ester, amide, phosphorylated, nitrosylated and succinate / seleno-succinate forms of tocotrienol.

[0083] Tocotrienols or derivatives thereof for use in the compositions and methods of the present invention can be extracted from natural sources or synthesized according to methods known in the art. Tocotrienols can be derived from plant extracts, such as palm oil, rice bran oil, malt, barley and annatto beans. In a preferred embodiment, tocotrienols are derived from palm oil or annatto.

[0084] The composition of the present invention may include one form of tocotrienol or its derivatives, or a mixture of different tocotrienols or their derivatives.

[0085] The tocotrienols in the composition of the present invention may be isolated from other components of the vitamin E extract, or may be present in combination with other vitamin E components.

[0086] In a preferred embodiment, the tocotrienol component of the pharmaceutical composition is greater than the tocopherol component.

[0087] Tocotrienols can be present in a tocotrienol-rich fraction produced from a vitamin E extract. The tocotrienol-rich fraction can include some alpha-tocopherol component from vitamin E. In a preferred embodiment, the tocotrienol-rich fraction includes no more than about 50%, preferably no more than about 40%, more preferably no more than about 30%, more preferably no more than about 20%, and more preferably no more than about 10% alpha-tocopherol.

[0088] In a preferred embodiment, the composition of the present invention comprises less than 10% tocopherol.

[0089] In another embodiment, the tocopherols present in the vitamin E extract can be removed, modified, so that the activity competing with the tocotrienols is eliminated or reduced. Those of ordinary skill in the art will appreciate that this can be achieved by many means, including but not limited to enzyme modification (see Torres et al. "Enzymatic Modification for Ascorbic Acid and Alpha-Tocopherol Enhances their Stability in Food and Nutritional Application" The Open Food Science Journal 2008, 2, 1-9).

[0090] In a preferred embodiment, the composition of the present invention comprises one or both of gamma-tocotrienol and delta-tocotrienol or their derivatives.

[0091] Transmucosal delivery refers to the route of administration of a drug through the mucous membranes. Transmucosal delivery includes within its scope sublingual; sublabial; buccal and nasal delivery.

[0092] Transmucosal delivery is achieved due to the connective tissue beneath the transmucosal epithelium containing capillaries that have the ability to allow some substances to diffuse through them and allow entry into the venous circulation.

[0093] The formulation for transmucosal delivery can be a solid dosage form (eg, tablet, capsule, lozenge or film), gel, liquid, emulsion, suspension, spray or aerosol formulation.

[0094] The formulation according to the present invention may include a pharmaceutically acceptable excipient or carrier suitable for a transmucosal delivery system. Suitable excipients are described in, for example, Handbook of Pharmacal Excipients, Rowe, Raymond C; Sheskey, Paul J; Cook, Walter G; Fenton, Marian E (editor), Pharmaceutical Press.

[0095] Suitable excipients include, but are not limited to, cyclodextrin, ethylcellulose, microcrystalline cellulose, cross-linked polyvinyl pyrrolidone, dicalcium phosphate, calcium carbonate, silicon dioxide, methylcellulose, carboxymethylcellulose, hydroxymethylcellulose, hydroxypropyl methylcellulose, alginates, gelatin, guar gum, tragacanth gum, gum arabic, polyacrylic acid, polymethacrylic acid, polysilicic acid and its salts, polylactic acid, polymaleic acid, polyvinyl alcohol, polyethylene glycol, polyvinyl pyrrolidone, nonionic block copolymers, carbomer, polycarbophil, polysorbate, and water-soluble starch.

[0096] Gel formulations may include the excipients carboxymethylcellulose and polyethylene glycol.

[0097] In a preferred embodiment of the present invention, the composition is formulated for sublingual delivery of at least one tocotrienol or a derivative thereof.

[0098] For purposes of the present invention, there are many examples of sublingual delivery methods and suitable carrier systems that can be used for the purpose of delivering tocotrienols, some of which can be found in "An Overview on: Sublingual Route for Systemic Drug Delivery" - International Journal of Research in Pharmaceutical and Biomedical Sciences Vol. 3 (2) April-June 2012, which is hereby incorporated by reference in its entirety.

[0099] Aspects of the invention include:

[0100] i) enhancing absorption of tocotrienols by formulation in a carrier system designed to facilitate absorption via transmucosal routes of administration including, but not limited to, oral, buccal, sublingual / sublabial, rectal, and nasal; and / or

[0101] ii) improving the delivery of tocotrienols to tissue targets, including through vesicular or nanoparticle formulations, such as self-assembling carrier systems that act like chylomicrons, low-density lipoproteins, cubosomes, mesophases, self-assembling amphiphiles, and non-self-assembling amphiphiles to increase the bioavailability of tocotrienols and minimize their hepatic metabolism; and / or

[0102] iii) Combination with other agents, such as plant extracts, vitamins (including but not limited to niacin) and minerals, proteins and lipids as well as carbohydrates and drugs that will give a synergistic effect when combined with tocotrienols.

[0103] Since tocotrienols have multiple effects, the balance between i) and ii) can be adjusted depending on the desired therapeutic effect. For example, if the natriuretic and diuretic effects of gamma-tocotrienol are the therapeutic target, the formulation will include gamma-tocotrienol alone or together with tocopherol, both of which are metabolized to 2,7,8-trimethyl-2-(b-carboxyethyl)-6-hydroxychroman (γ-CEHC), which acts on the kidneys to increase sodium excretion.

[0104] It is increasingly recognized that it is important to reduce alpha-tocopherol in the diet or in supplemental intake to reduce interference with the cholesterol-lowering effects of tocotrienols, especially gamma- and delta-tocotrienols. A study conducted in chickens revealed that the reduction in lipid parameters, including cholesterol production, was greater in birds given the least amount of alpha-tocopherol relative to tocotrienols. In contrast, birds given the highest amount of alpha-tocopherol had increased cholesterol production (Qureshi et al. "Dietary alpha-tocopherol attenuates the impact of gamma-tocotrienol on hepatic 3-hydroxy-3-methylglutaryl coenzyme A reductase activity in chickens" J Nutrition 1996 Feb; 126(2): 389-94). A separate study demonstrated that high levels of alpha-tocopherol increased cholesterol production (Khor et al. "Lipidaemic effects of tocotrienols, tocopherols and squalene: studies in the hamster," Asia Pacific J Clin Nutr (1997) 6(1):36-40). Interference of alpha-tocopherol with tocotrienol absorption has also been previously described by Ikeda et al., who showed that alpha-tocopherol interferes with the absorption of alpha-tocotrienol but not gamma-tocotrienol (Ikeda et al. "Dietary alpha-tocopherol decreases alpha tocotrienol but not gamma-tocotrienol concentration in rats" J Nutr 2003 Feb;133(2):428-34).Recently, Japanese researchers have found that tocopherols, and in particular alpha-tocopherol, interfere with the ability of delta-tocotrienol to induce apoptosis in cancer cells, while blocking the absorption of delta-tocotrienol (Shibata et al. "alpha-Tocopherol attenuates the cytotoxic effect of delta-tocotrienol in human colorectal adenocarcinoma cells" Biocehm. Piophys. Res. Commun. 397(2):214-9). Finally, alpha-tocopherol was shown to interfere with tocotrienols by increasing their catabolism (Sontag et al. "Influence of major structural features of tocopherols and tocotrienols on their omega oxidation by tocopherol-omega hydroxylase" J. Lipid Res. 48(5): 1090-8).

[0105] It is to be understood that therapeutic agents comprising one or more tocotrienols or derivatives thereof are included within the scope of the present invention and that these therapeutic agents:

[0106] (a) does not contain tocopherol;

[0107] (b) contains tocopherol levels which have been minimized as far as possible; or

[0108] (c) comprises tocopherols which have been modified so as to ensure that any interference of the tocopherols with the tocotrienols is reduced or eliminated.

[0109] Similarly, include the combination therapy that can produce synergistic effect with tocotrienol.For example, using i) and iii), γ or δ-tocotrienol can be combined in one or more self-assembly carrier systems designed for sublingual absorption with a kind of beta-drug (itself is amphipathic).This can bypass the liver metabolism of tocotrienol and avoid one of the key side effects of beta-drug, gastrointestinal discomfort.The combined effect can then reduce biosynthetic cholesterol (the minimizing of HMG-CoA reductase) by tocotrienol and increase the HDL+ in triglyceride by beta-drug (acting via PPAR-α).

[0110] The tocotrienols in the compositions of the present invention can be associated with amphiphilic chemicals (including those that can self-assemble into molecular structures, such as carbosomes and mesophases) that enhance sublingual absorption. Specifically, tocotrienols can be associated with self-assembling amphiphilic chemicals that can assemble into structures called carbosomes. The assembly of tocotrienols into carbosomes can also inhibit the metabolic degradation of tocotrienols. Carbosomes are bicontinuous cubic liquid crystal materials that allow the simultaneous incorporation of water-soluble materials and oil-soluble materials and amphiphilic molecules. The use of carbosomes can enhance the bioavailability of tocotrienols and allow tocotrienols to bypass the liver, which can interfere with the delivery of tocotrienols.

[0111] Carbosomes can be characterized as having a high internal surface area along with a cubic crystal structure and also include advantages like simple preparation process, biodegradability of lipids, ability to encapsulate hydrophobic, hydrophilic, amphiphilic substances, targeted bioactive agents along with their sustained release. The preparation of carbosomes typically involves emulsification of monoglycerides along with polymers, followed by sonication and homogenization. Alternative preparation methods of carbosomes known in the art may also be utilized.

[0112] The compositions of the present invention may also include solid lipid core nanoparticles in the formulation, which act as chylomicrons to enhance the absorption of lipids from the intestine into the lymphatic system, thereby bypassing the liver (Paliwal, R et al. "Effect of lipid core material on characteristics of solid lipid nanoparticles designed for oral lymphatic delivery", Nanomedicine: Nanotechnology, Biology and Medicine 5 (2009) 184-191).

[0113] The scope of the present invention also encompasses carrier systems such as synthetic nano-low-density lipoproteins (Nikanjam, 2006). Such systems allow the delivery of lipophilic drugs via the LDL-receptors of cells, thereby enhancing absorption into body tissues without the need for repackaging into natural lipoproteins and export through the liver.

[0114] In one embodiment of the present invention, tocotrienol or its derivatives may be formulated as a sublingual or buccal dosage form adapted from the formulation described in Australian Patent Application No. 2010262738 (Cumming et al.).

[0115] In another embodiment, the composition of the present invention includes at least one tocotrienol or its derivatives formulated as vesicles or particles, such as liposomes, vesicles, transfer bodies, pharmacoplasts and nanoparticles. Such vesicles and particles include, but are not limited to, those described in the following: Gangwar, M. et al. "Recent Advances in various emerging vesicular systems: An overview", Asia Pacific Journal of Tropical Biomedicine (2012) S1176-S1188, Paliwal, R et al. (2009), Sagnella, SM et al. "Anandamide and analogous endocannabinoids: a lipid self-assembly study", Soft Matter, 2011, 7, 5319, and Hood, E. et al. "Nanocarriers for vascular delivery of antioxidants", Nanomedicine (Lond) Sep 2011; 6(7): 1257-1272, and Gupta, S. et al. "Polyether based amphiphiles for delivery of active components", Polymer 53 (2012) 3053-3078, which are hereby incorporated by reference in their entirety.

[0116] The transmucosal tocotrienol compositions of the present invention can be produced by the method described in WO 2012 / 109694 (Ko. Sai, Ying), which is hereby incorporated by reference in its entirety. From the disclosure of Ko, the water-soluble components in the disclosed compositions and formulations dissolve when in contact with saliva, resulting in the formation of micropores on the coating surface, which promote the disintegration of the microcapsules into the liquid gel. The gel is arranged along the mucosal surface, thereby delivering the active ingredient and optimizing absorption. In addition, Ko shows that this sublingual delivery platform can transport molecules across the sublingual mucosa, such as proteins that include both hydrophilic and hydrophobic parts.

[0117] The compositions of the present invention may include tocotrienol, β-cyclodextrin (and / or other cyclodextrin variants) in an appropriate ratio so as to retain the tocotrienol within the hydrophobic core of the cyclodextrin molecule. An example of such a formulation is found in the work of Mannila J, Jarvinen T, Jarvine K & Jarho P, "Precipitation complexation method produces cannabidiol / beta cyclodextrin inclusion complex suitable for sublingual administration on cannabidiol" Journal of Pharmaceutical Sciences 96, 312-319, 2007. Tocotrienols are known to form complexes with gamma-cyclodextrin, but this formulation is not suitable due to the degradation of gamma-cyclodextrin by salivary amylase, whereas the alpha and beta forms are resistant (Ikeda S, Uchida T, Ichikawa T et al., "Complexation of tocotrienol with gamma-cyclodextrin enhances intestinal absorption of tocotrienols Complexation of tocotrienols with gamma-cyclodextrin enhances intestinal absorption of tocotrienols in rats” Bioscience, Biotechnology and Biochemistry 74, 1453-1457, 2010).

[0118] A wide variety of formulations and compositions may be utilized for accomplishing the purposes of the present invention, provided that the compositions and formulations include pharmaceutically acceptable excipients and carriers which, when exposed to an aqueous / salivary mucosal environment, will allow the tocotrienols present in the composition to remain in contact with the mucosal surface for a sufficient period of time to allow absorption, promote or at least not hinder absorption of the tocotrienols through the mucosa and allow the tocotrienols to be absorbed in a bioavailable form.

[0119] In another embodiment, transmucosally administered tocotrienols can be combined with any other compounds that will complement and enhance the therapeutic effects of tocotrienols or their derivatives, including but not limited to monoglycerides, lignans, isoprenoids, amino acids, CoQ10, polyphenols, omega-3 fatty acids, endocannabinoid system agonists and antagonists, flavonoids, carotenoids, mono- and oligosaccharides, niacin, and bioactive peptides.

[0120] In another embodiment, transmucosally administered tocotrienols may be combined with an extract from sesame seeds and or sesame lignans.

[0121] It should also be understood that derivatives of tocotrienols that enhance the therapeutic effect are also included, such as phosphorylated, nitrosylated, and succinate / seleno-succinate forms. An example of a method for modifying tocotrienols can be found in Vraka et al. "Synthesis and study of the cancer cell growth inhibitory properties of α, γ-tocopheryl and γ-tocotrienyl 2-phenylselenyl succinates" Bioorganic & Medicinal Chemistry 14 (2006) 2684-2696, which is hereby incorporated by reference in its entirety.

[0122] In a further embodiment, tocotrienols (and / or their derivatives) are administered in combination with an additional compound designed to complement and enhance the therapeutic effect of tocotrienols or their derivatives, selected from the group consisting of monoglycerides; lignans; isoprenoids; polyphenols; flavonoids; carotenoids; mono- and oligosaccharides; niacin and bioactive peptides in the form of a pharmaceutical composition suitable for buccal, sublingual, mucosal or nasal administration, but preferably sublingual administration.

[0123] In another embodiment, tocotrienols (and / or derivatives thereof) are administered in the form of a pharmaceutical composition suitable for buccal, sublingual, mucosal or nasal administration, preferably sublingual administration, and wherein tocopherols are absent, or if tocopherols are present, the interfering and / or competing activity of any tocopherols present in the pharmaceutical composition (relative to tocotrienols) has been eliminated, reduced or minimized. The composition may include or be administered in combination with any other compound that will complement and enhance the therapeutic effect of tocotrienols, such as monoglycerides, lignans, isoprenoids, polyphenols, flavonoids, carotenoids, mono- and oligosaccharides, niacin, and bioactive peptides.

[0124] Orally ingested tocotrienols have lower bioavailability than orally ingested tocopherols, as shown by reduced peak concentrations in plasma and shorter half-lives after ingestion in a recent human study (S, Fairus, S. et al. Nutr J. 2012 Jan 17; 11:5). An animal study has estimated that the oral bioavailability of tocotrienols is 4-fold lower than that of tocopherols (9% vs. 36%) (Abuasal, BS. et al. Biopharm Drug Dispos. 2012 Jul; 33(5): 246-56). The mechanisms that determine the bioavailability of tocotrienols have not been fully elucidated, but intestinal absorption and liver processing appear to be key factors.

[0125] The absorption of vitamin E isomers varies between individuals, possibly due to differences in the expression of different transporters (e.g., scavenger receptor B1 and Niemann-Pick C1L1 receptors), as well as the presence of other lipids (e.g., cholesterol) that share these transport mechanisms (Reboul, E. and Borel, P. Prog Lipid Res 2011 50:388-402). Hepatic processing is another source of biological distinction between tocopherol and tocotrienol isomers, based on differences in affinity for the intracellular alpha-tocopherol transporter (a-TTP). The relative affinity of vitamin E isomers for a-TTP has been demonstrated to be in the following order: alpha-tocopherol (100%) > alpha-tocotrienol (12%) > gamma-tocotrienol (9%) > delta-tocopherol (2%) (Hosomi, A. et al. FEBS Letters 1997, 409: 105-108). This mechanism helps explain the presence of alpha-tocopherol as the major vitamin E isomer detected in triglyceride-rich particles, low-density and high-density lipoproteins, and the rapid disappearance of alpha-tocotrienol, gamma-tocotrienol, and delta-tocotrienol from circulating plasma and lipoproteins. In humans, the plasma concentration of α-tocotrienol is 2-fold higher than that of γ-tocotrienol and almost 10-fold higher than that of δ-tocotrienol after supplementation with the same dose of tocotrienol preparations (O'Byrne, D. et al. Free Radic Biol Med 2000, 29:834-845).

[0126] Without wishing to be bound by theory, it is believed that sublingual delivery of tocotrienols or derivatives thereof according to the present invention has at least two distinct advantages over conventional oral delivery. First, it can overcome the problem of low intestinal absorption of tocotrienols by ensuring high absorption via the sublingual lymphatic system. This technology will also bypass the low affinity of tocotrienols for the alpha-tocopherol transporter, because the lymphatic circulation will deliver tocotrienols to target tissues without the need for incorporation into triglycerides and lipoproteins by the liver and export to the circulation. This approach will also minimize the first-pass metabolism of tocotrienols by the liver and increase the amount of tocotrienols delivered to other organs.

[0127] In addition, by using sublingual delivery systems in combination with other technologies, such as assembling tocotrienols into nanoparticles, in their native form or as chemical derivatives, or in combination with other molecules (Abuasal, BS et al. Lipids. 2012 May;47(5):461-9. doi:10.1007 / s 11745-012-3655-4. Epub 2012 Jan 24), delivery and bioactivity can be tuned to specific target tissues and therapeutic applications.

[0128] Therefore, in a further embodiment, the composition of the present invention comprises tocotrienol or a derivative thereof assembled into nanoparticles.

[0129] The inventors of the present application have confirmed a previous study (Lee, SP, Mar, GY & Ng, LT 2009, European Journal of Applied Physiology, 107, 587-595) that tocotrienols can improve exercise endurance in normal weight Wistar rats by showing that this improvement also extends to Sprague-Dawley rats with diet-induced obesity. In addition, the inventors of the present application have demonstrated an unexpected increase in efficacy of sublingual formulations of tocotrienols over oral formulations in improving exercise endurance in humans.

[0130] Thus, in another aspect, the present invention provides a method for improving exercise endurance and performance in animals, and particularly humans, by administering tocotrienols (and / or derivatives thereof) in the form of a pharmaceutical composition suitable for buccal, sublingual, mucosal or nasal administration, preferably sublingual administration.

[0131] In another aspect, the present invention provides a method for improving exercise endurance and behavioral performance in animals, and particularly humans, by administering tocotrienols (and / or derivatives thereof) in the form of a pharmaceutical composition suitable for buccal, sublingual, mucosal or nasal administration, preferably sublingual administration, in combination with any other compounds that will complement and enhance the improving effects of tocotrienols with respect to exercise endurance, such as monoglycerides, lignans, isoprenoids, polyphenols, flavonoids, carotenoids, mono- and oligosaccharides, niacin and bioactive peptides.

[0132] In another aspect, the present invention provides a method of improving exercise endurance and performance in animals, and particularly humans, by administering tocotrienols (and / or derivatives thereof) in the form of a pharmaceutical composition suitable for buccal, sublingual, mucosal or nasal administration, but preferably sublingual administration, and wherein the tocotrienol component is greater than the tocopherol component and / or tocopherol is absent, or if tocopherol is present, the interfering and / or competing activity of any tocopherol present in the pharmaceutical composition has been eliminated, reduced or minimized. The composition may include or be administered in combination with any other compound that will complement and enhance the improving effect of tocotrienols on exercise endurance, such as monoglycerides, lignans, isoprenoids, polyphenols, flavonoids, carotenoids, mono- and oligosaccharides, niacin and bioactive peptides.

[0133] In another aspect, the present invention provides a method of promoting weight loss in animals, and particularly humans, by administering tocotrienols (and / or derivatives thereof) in the form of a pharmaceutical composition suitable for buccal, sublingual, mucosal or nasal administration, but preferably sublingual administration.

[0134] In another aspect, the present invention provides a method of promoting weight loss in animals, and particularly humans, by administering tocotrienols (and / or derivatives thereof) in the form of a pharmaceutical composition suitable for buccal, sublingual, mucosal or nasal administration, but preferably sublingual administration, in combination with any other compounds that will complement and enhance the effects of tocotrienols with respect to promoting weight loss, such as monoglycerides, lignans, isoprenoids, polyphenols, flavonoids, carotenoids, mono- and oligosaccharides, niacin and bioactive peptides.

[0135] In another aspect, the present invention provides a method of promoting weight loss in animals, and particularly humans, by administering tocotrienols (and / or derivatives thereof) in the form of a pharmaceutical composition suitable for buccal, sublingual, mucosal or nasal administration, but preferably sublingual administration, and wherein the tocotrienol component is greater than the tocopherol component, or tocopherol is absent, or if tocopherol is present, the interfering and / or competing activity of any tocopherol present in the pharmaceutical composition has been eliminated, reduced or minimized. The composition may include, or be administered in combination with, any other compounds that will complement and enhance the effects of tocotrienols with respect to promoting weight loss, such as monoglycerides, lignans, isoprenoids, polyphenols, flavonoids, carotenoids, mono- and oligosaccharides, niacin, and bioactive peptides.

[0136] Tocotrienols have been shown to improve glycemic control and insulin response (Kuhad et al. (2009)).

[0137] Thus, in another aspect, the present invention provides a method for stabilizing and / or controlling blood glucose levels in animals, and particularly humans, by administering tocotrienols (and / or derivatives thereof) in the form of a pharmaceutical composition suitable for buccal, sublingual, mucosal or nasal administration, but preferably sublingual administration.

[0138] In another aspect, the present invention provides a method for stabilizing and / or controlling blood glucose levels in animals, and particularly humans, by administering tocotrienols (and / or derivatives thereof) in the form of a pharmaceutical composition suitable for buccal, sublingual, mucosal or nasal administration, but preferably sublingual administration, in combination with any other compounds that will complement and enhance the effects of tocotrienols with respect to stabilizing and / or controlling blood glucose levels, such as monoglycerides, lignans, isoprenoids, polyphenols, flavonoids, carotenoids, mono- and oligosaccharides, niacin and bioactive peptides.

[0139] In another aspect, the present invention provides a method of stabilizing and / or controlling blood glucose levels in animals, and particularly humans, by administering tocotrienols (and / or derivatives thereof) in the form of a pharmaceutical composition suitable for buccal, sublingual, mucosal or nasal administration, but preferably sublingual administration, and wherein the tocotrienol component is greater than the tocopherol component, or tocopherol is absent, or if tocopherol is present, the interfering and / or competing activity of any tocopherol present in the pharmaceutical composition has been eliminated, reduced or minimized. The composition may include, or be administered in combination with, any other compound that will complement and enhance the effects of tocotrienols with respect to stabilizing and / or controlling blood glucose levels, such as monoglycerides, lignans, isoprenoids, polyphenols, flavonoids, carotenoids, mono- and oligosaccharides, niacin and bioactive peptides.

[0140] In another aspect, the present invention provides a method of reducing hypertension in animals, and particularly humans, by administering tocotrienols (and / or derivatives thereof) in the form of a pharmaceutical composition suitable for buccal, sublingual, transdermal, oral, mucosal or nasal administration, but preferably sublingual administration.

[0141] In another aspect, the present invention provides a method of reducing hypertension in animals, and particularly humans, by administering tocotrienols (and / or derivatives thereof) in the form of a pharmaceutical composition suitable for buccal, sublingual, mucosal or nasal administration, but preferably sublingual administration, in combination with any other compounds that will complement and enhance the effects of tocotrienols with respect to reducing hypertension, such as monoglycerides, lignans, isoprenoids, polyphenols, flavonoids, carotenoids, mono- and oligosaccharides, niacin and bioactive peptides.

[0142] In another aspect, the present invention provides a method of reducing hypertension in animals, and particularly humans, by administering tocotrienols (and / or derivatives thereof) in the form of a pharmaceutical composition suitable for buccal, sublingual, mucosal or nasal administration, but preferably sublingual administration, and wherein the tocotrienol component is greater than the tocopherol component, or tocopherol is absent, or if tocopherol is present, the interfering and / or competing activity of any tocopherol present in the pharmaceutical composition has been eliminated, reduced or minimized. The composition may include, or be administered in combination with, any other compound that will complement and enhance the effects of tocotrienols with respect to reducing hypertension, such as monoglycerides, lignans, isoprenoids, polyphenols, flavonoids, carotenoids, mono- and oligosaccharides, niacin and bioactive peptides.

[0143] In another aspect, the present invention provides a method for treating ischemic diseases in animals, and particularly humans, by administering tocotrienols (and / or derivatives thereof) in the form of a pharmaceutical composition suitable for buccal, sublingual, mucosal or nasal administration, but preferably sublingual administration.

[0144] In another aspect, the present invention provides a method of treating ischemic diseases in animals, and particularly humans, by administering tocotrienols (and / or derivatives thereof) in the form of a pharmaceutical composition suitable for buccal, sublingual, mucosal or nasal administration, but preferably sublingual administration, in combination with any other compounds that will supplement and enhance the effects of tocotrienols with respect to treating ischemic diseases, such as monoglycerides, lignans, isoprenoids, polyphenols, flavonoids, carotenoids, mono- and oligosaccharides, niacin and bioactive peptides.

[0145] In another aspect, the present invention provides a method of treating ischemic disease in animals, and particularly humans, by administering tocotrienols (and / or derivatives thereof) in the form of a pharmaceutical composition suitable for buccal, sublingual, mucosal or nasal administration, but preferably sublingual administration, and wherein the tocotrienol component is greater than the tocopherol component, or tocopherol is absent, or if tocopherol is present, the interfering and / or competing activity of any tocopherol present in the pharmaceutical composition has been eliminated, reduced or minimized. The composition may include, or be administered in combination with, any other compound that will complement and enhance the effects of tocotrienols with respect to treating ischemic disease, such as monoglycerides, lignans, isoprenoids, polyphenols, flavonoids, carotenoids, mono- and oligosaccharides, niacin, and bioactive peptides.

[0146] In another aspect, the present invention provides a method for reducing cholesterol and / or triglycerides in animals, and particularly humans, by administering tocotrienols (and / or derivatives thereof) in the form of a pharmaceutical composition suitable for buccal, sublingual, mucosal or nasal administration, but preferably sublingual administration.

[0147] In another aspect, the present invention provides a method for reducing cholesterol and / or triglycerides in animals, and particularly humans, by administering tocotrienols (and / or derivatives thereof) in the form of a pharmaceutical composition suitable for buccal, sublingual, mucosal or nasal administration, but preferably sublingual administration, in combination with any other compounds that will complement and enhance the effects of tocotrienols with respect to reducing cholesterol and / or triglycerides, such as monoglycerides, lignans, isoprenoids, polyphenols, flavonoids, carotenoids, mono- and oligosaccharides, niacin and bioactive peptides.

[0148] In another aspect, the present invention provides a method of reducing cholesterol and / or triglycerides in animals, and particularly humans, by administering tocotrienols (and / or derivatives thereof) in the form of a pharmaceutical composition suitable for buccal, sublingual, mucosal or nasal administration, but preferably sublingual administration, and wherein the tocotrienol component is greater than the tocopherol component, or tocopherol is absent, or if tocopherol is present, the interfering and / or competing activity of any tocopherol present in the pharmaceutical composition has been eliminated, reduced or minimized. The composition may include, or be administered in combination with, any other compound that will complement and enhance the effect of tocotrienols with respect to reducing cholesterol and / or triglycerides, such as monoglycerides, lignans, isoprenoids, polyphenols, flavonoids, carotenoids, mono- and oligosaccharides, niacin and bioactive peptides.

[0149] In another aspect, the present invention provides a method of treating cancer in animals, and particularly humans, by administering tocotrienols (and / or derivatives thereof) in the form of a pharmaceutical composition suitable for buccal, sublingual, transdermal, oral, mucosal or nasal administration, but preferably sublingual administration.

[0150] In another aspect, the present invention provides a method of treating cancer in animals, and particularly humans, by administering tocotrienols (and / or derivatives thereof) in the form of a pharmaceutical composition suitable for buccal, sublingual, mucosal or nasal administration, but preferably sublingual administration, in combination with any other compounds that will complement and enhance the effects of tocotrienols with respect to treating cancer, such as monoglycerides, lignans, isoprenoids, polyphenols, flavonoids, carotenoids, mono- and oligosaccharides, niacin and bioactive peptides.

[0151] In another aspect, the present invention provides a method of treating cancer in animals, and particularly humans, by administering tocotrienols (and / or derivatives thereof) in the form of a pharmaceutical composition suitable for buccal, sublingual, mucosal or nasal administration, but preferably sublingual administration, and wherein the tocotrienol component is greater than the tocopherol component, or tocopherol is absent, or if tocopherol is present, the interfering and / or competing activity of any tocopherol present in the pharmaceutical composition has been eliminated, reduced or minimized. The composition may include, or be administered in combination with, any other compound that will complement and enhance the effects of tocotrienols with respect to treating cancer, such as monoglycerides, lignans, isoprenoids, polyphenols, flavonoids, carotenoids, mono- and oligosaccharides, niacin, and bioactive peptides.

[0152] In another aspect, the present invention provides a method of treating cancer in animals, and particularly humans, by administering tocotrienols (and / or derivatives thereof) in the form of a pharmaceutical composition suitable for buccal, sublingual, mucosal or nasal administration, but preferably sublingual administration.

[0153] In another aspect, the present invention provides a method of increasing the bioavailability of tocotrienols administered to animals, and particularly humans, by administering tocotrienols (and / or derivatives thereof) in the form of a pharmaceutical composition suitable for buccal, sublingual, mucosal or nasal administration, but preferably sublingual administration, in combination with any other compounds that will complement and enhance the effects of tocotrienols with respect to increasing the bioavailability of tocotrienols administered to the animal, such as monoglycerides, lignans, isoprenoids, polyphenols, flavonoids, carotenoids, mono- and oligosaccharides, niacin and bioactive peptides.

[0154] In another aspect, the present invention provides a method of increasing the bioavailability of tocotrienols administered to animals, and particularly humans, by administering tocotrienols (and / or derivatives thereof) in the form of a pharmaceutical composition suitable for buccal, sublingual, mucosal or nasal administration, but preferably sublingual administration, and wherein the tocotrienol component is greater than the tocopherol component, tocopherols are absent, or if tocopherols are present, the interfering and / or competing activity of any tocopherols present in the pharmaceutical composition has been eliminated, reduced or minimized. The composition may include or be administered in combination with any other compound that will complement and enhance the effect of tocotrienols with respect to increasing the bioavailability of tocotrienols administered to animals, such as monoglycerides, lignans, isoprenoids, polyphenols, flavonoids, carotenoids, mono- and oligosaccharides, niacin, and bioactive peptides.

[0155] In another aspect, the present invention provides a method for minimizing the dosage required to achieve a therapeutic effect by administering tocotrienols to animals, and particularly humans, by administering tocotrienols (and / or derivatives thereof) in the form of a pharmaceutical composition suitable for buccal, sublingual, mucosal or nasal administration, but preferably sublingual administration.

[0156] In another aspect, the present invention provides a method of minimizing the dose required to achieve a therapeutic effect by administering tocotrienols to animals, and particularly humans, by administering tocotrienols (and / or derivatives thereof) in the form of a pharmaceutical composition suitable for buccal, sublingual, mucosal or nasal administration, but preferably sublingual administration, in combination with any other compounds that will complement and enhance the effect of tocotrienols with respect to minimizing the dose required to achieve a therapeutic effect by administering tocotrienols, such as monoglycerides, lignans, isoprenoids, polyphenols, flavonoids, carotenoids, mono- and oligosaccharides, niacin and bioactive peptides.

[0157] In another aspect, the present invention provides a method for minimizing the dose required to achieve a therapeutic effect by administering tocotrienols to animals, and particularly humans, by administering tocotrienols (and / or derivatives thereof) in a pharmaceutical composition suitable for buccal, sublingual, mucosal or nasal administration, but preferably sublingual administration, and wherein the tocotrienol component is greater than the tocopherol component, or tocopherol is absent, or if tocopherol is present, the interfering and / or competing activity of any tocopherol present in the pharmaceutical composition has been eliminated, reduced or minimized. The composition may include or be administered in combination with any other compound that will complement and enhance the effect of tocotrienols with respect to minimizing the dose required to achieve a therapeutic effect by administering tocotrienols, such as monoglycerides, lignans, isoprenoids, polyphenols, flavonoids, carotenoids, mono- and oligosaccharides, niacin, and bioactive peptides.

[0158] In another aspect, the present invention provides a method of reducing and / or inhibiting inflammation by administering tocotrienols to animals, and particularly humans, by administering tocotrienols (and / or derivatives thereof) in the form of a pharmaceutical composition suitable for buccal, sublingual, mucosal or nasal administration, but preferably sublingual administration.

[0159] In another aspect, the present invention provides a method of reducing or inhibiting inflammation by administering tocotrienols to animals, and particularly humans, by administering tocotrienols (and / or derivatives thereof) in the form of a pharmaceutical composition suitable for buccal, sublingual, mucosal or nasal administration, but preferably sublingual administration, in combination with any other compounds that will complement and enhance the effects of tocotrienols with respect to reducing and / or inhibiting inflammation by administration of tocotrienols, such as monoglycerides, lignans, isoprenoids, polyphenols, flavonoids, carotenoids, mono- and oligosaccharides, niacin and bioactive peptides.

[0160] In another aspect, the present invention provides a method of reducing and / or inhibiting inflammation by administering tocotrienols to animals, and particularly humans, by administering tocotrienols (and / or derivatives thereof) in the form of a pharmaceutical composition suitable for buccal, sublingual, mucosal or nasal administration, but preferably sublingual administration, and wherein the tocotrienol component is greater than the tocopherol component, or tocopherols are absent, or if tocopherols are present, the interfering and / or competing activity of any tocopherols present in the pharmaceutical composition has been eliminated, reduced or minimized. The composition may include or be administered in combination with any other compound that will complement and enhance the effect of tocotrienols with respect to reducing and / or inhibiting inflammation by administering tocotrienols, such as monoglycerides, lignans, isoprenoids, polyphenols, flavonoids, carotenoids, mono- and oligosaccharides, niacin, and bioactive peptides.

[0161] In another aspect, the present invention provides a method of reducing and / or inhibiting post-exercise muscle soreness by administering tocotrienols to animals, and particularly humans, by administering tocotrienols (and / or derivatives thereof) in the form of a pharmaceutical composition suitable for buccal, sublingual, mucosal or nasal administration, but preferably sublingual administration.

[0162] In another aspect, the present invention provides a method for reducing and / or inhibiting post-exercise muscle soreness by administering tocotrienols to animals, and particularly humans, by administering tocotrienols (and / or derivatives thereof) in the form of a pharmaceutical composition suitable for buccal, sublingual, mucosal or nasal administration, but preferably sublingual administration, in combination with any other compounds that will complement and enhance the effects of tocotrienols with respect to reducing and / or inhibiting post-exercise muscle soreness by administration of tocotrienols, such as monoglycerides, lignans, isoprenoids, polyphenols, flavonoids, carotenoids, mono- and oligosaccharides, niacin and bioactive peptides.

[0163] In another aspect, the present invention provides a method of reducing and / or inhibiting post-exercise muscle soreness by administering tocotrienols (and / or derivatives thereof) to animals, and particularly humans, by administering tocotrienols (and / or derivatives thereof) in the form of a pharmaceutical composition suitable for buccal, sublingual, mucosal or nasal administration, but preferably sublingual administration, and wherein the tocotrienol component is greater than the tocopherol component, or tocopherols are absent, or if tocopherols are present, the interfering and / or competing activity of any tocopherols present in the pharmaceutical composition has been eliminated, reduced or minimized. The composition may include or be administered in combination with any other compound that will complement and enhance the effect of tocotrienols with respect to reducing and / or inhibiting post-exercise muscle soreness by administering tocotrienols, such as monoglycerides, lignans, isoprenoids, polyphenols, flavonoids, carotenoids, mono- and oligosaccharides, niacin, and bioactive peptides.

[0164] In another aspect, the present invention provides a method of reducing and / or inhibiting delayed onset muscle soreness by administering tocotrienols to animals, and particularly humans, by administering tocotrienols (and / or derivatives thereof) in the form of a pharmaceutical composition suitable for buccal, sublingual, mucosal or nasal administration, but preferably sublingual administration.

[0165] In another aspect, the present invention provides a method of reducing and / or inhibiting delayed onset muscle soreness by administering tocotrienols to animals, and particularly humans, by administering tocotrienols (and / or derivatives thereof) in the form of a pharmaceutical composition suitable for buccal, sublingual, mucosal or nasal administration, but preferably sublingual administration, in combination with any other compounds that will complement and enhance the effects of tocotrienols with respect to reducing and / or inhibiting delayed onset muscle soreness by administration of tocotrienols, such as monoglycerides, lignans, isoprenoids, polyphenols, flavonoids, carotenoids, mono- and oligosaccharides, niacin and bioactive peptides.

[0166] In another aspect, the present invention provides a method of reducing and / or inhibiting delayed onset muscle soreness by administering tocotrienols (and / or derivatives thereof) to animals, and particularly humans, by administering tocotrienols (and / or derivatives thereof) in the form of a pharmaceutical composition suitable for buccal, sublingual, mucosal or nasal administration, but preferably sublingual administration, and wherein the tocotrienol component is greater than the tocopherol component, or tocopherols are absent, or if tocopherols are present, the interfering and / or competing activity of any tocopherols present in the pharmaceutical composition has been eliminated, reduced or minimized. The composition may include or be administered in combination with any other compound that will complement and enhance the effect of tocotrienols with respect to reducing and / or inhibiting delayed onset muscle soreness by administering tocotrienols, such as monoglycerides, lignans, isoprenoids, polyphenols, flavonoids, carotenoids, mono- and oligosaccharides, niacin, and bioactive peptides.

[0167] In another aspect, the present invention provides a method of reducing and / or treating myocardial fibrosis by administering tocotrienols to animals, and particularly humans, by administering tocotrienols (and / or derivatives thereof) in the form of a pharmaceutical composition suitable for buccal, sublingual, mucosal or nasal administration, but preferably sublingual administration.

[0168] In another aspect, the present invention provides a method for reducing and / or treating myocardial fibrosis by administering tocotrienols to animals, and particularly humans, by administering tocotrienols (and / or derivatives thereof) in the form of a pharmaceutical composition suitable for buccal, sublingual, mucosal or nasal administration, but preferably sublingual administration, in combination with any other compounds that will complement and enhance the effects of tocotrienols with respect to reducing and / or treating myocardial fibrosis by administration of tocotrienols, such as monoglycerides, lignans, isoprenoids, polyphenols, flavonoids, carotenoids, mono- and oligosaccharides, niacin and bioactive peptides.

[0169] In another aspect, the present invention provides a method of reducing and / or treating myocardial fibrosis by administering tocotrienols (and / or derivatives thereof) to animals, and particularly humans, in the form of a pharmaceutical composition suitable for buccal, sublingual, mucosal or nasal administration, but preferably sublingual administration, and wherein the tocotrienol component is greater than the tocopherol component, or tocopherol is absent, or if tocopherol is present, the interfering and / or competing activity of any tocopherol present in the pharmaceutical composition has been eliminated, reduced or minimized. The composition may include or be administered in combination with any other compound that will complement and enhance the effect of tocotrienols with respect to reducing and / or treating myocardial fibrosis by administering tocotrienols, such as monoglycerides, lignans, isoprenoids, polyphenols, flavonoids, carotenoids, mono- and oligosaccharides, niacin, and bioactive peptides.

[0170] In another aspect, the present invention provides a method of treating radiation exposure in animals by administering tocotrienols (and / or derivatives thereof) to animals, and particularly humans, by administering tocotrienols in the form of a pharmaceutical composition suitable for buccal, sublingual, mucosal or nasal administration, but preferably sublingual administration.

[0171] In another aspect, the present invention provides a method of treating radiation exposure in animals by administering tocotrienols to animals, and particularly humans, by administering tocotrienols (and / or derivatives thereof) in the form of a pharmaceutical composition suitable for buccal, sublingual, mucosal or nasal administration, but preferably sublingual administration, in combination with any other compounds that will complement and enhance the effects of tocotrienols with respect to treating radiation exposure in animals by administering tocotrienols, such as monoglycerides, lignans, isoprenoids, polyphenols, flavonoids, carotenoids, mono- and oligosaccharides, niacin and bioactive peptides.

[0172] In another aspect, the present invention provides a method of treating radiation exposure in animals by administering tocotrienols (and / or derivatives thereof) to animals, and particularly humans, in the form of a pharmaceutical composition suitable for buccal, sublingual, mucosal or nasal administration, but preferably sublingual administration, and wherein the tocotrienol component is greater than the tocopherol component, or tocopherol is absent, or if tocopherol is present, the interfering and / or competing activity of any tocopherol present in the pharmaceutical composition has been eliminated, reduced or minimized. The composition may include or be administered in combination with any other compound that will complement and enhance the effects of tocotrienols with respect to treating radiation exposure in animals by administering tocotrienols, such as monoglycerides, lignans, isoprenoids, polyphenols, flavonoids, carotenoids, mono- and oligosaccharides, niacin, and bioactive peptides.

[0173] In another aspect, the present invention provides a method of treating androgenetic alopecia by administering tocotrienols (and / or derivatives thereof) to animals, and particularly humans, by administering tocotrienols in the form of a pharmaceutical composition suitable for buccal, sublingual, mucosal or nasal administration, but preferably sublingual administration.

[0174] In another aspect, the present invention provides a method of treating androgenetic alopecia by administering tocotrienols to animals, and particularly humans, by administering tocotrienols (and / or derivatives thereof) in the form of a pharmaceutical composition suitable for buccal, sublingual, mucosal or nasal administration, but preferably sublingual administration, in combination with any other compounds that will complement and enhance the effects of tocotrienols with respect to the treatment of androgenetic alopecia by the administration of tocotrienols, such as monoglycerides, lignans, isoprenoids, polyphenols, flavonoids, carotenoids, mono- and oligosaccharides, niacin and bioactive peptides.

[0175] In another aspect, the present invention provides a method of treating androgenetic alopecia by administering tocotrienols (and / or derivatives thereof) to animals, and particularly humans, by administering tocotrienols (and / or derivatives thereof) in the form of a pharmaceutical composition suitable for buccal, sublingual, mucosal or nasal administration, but preferably sublingual administration, and wherein the tocotrienol component is greater than the tocopherol component, or tocopherols are absent, or if tocopherols are present, the interfering and / or competing activity of any tocopherols present in the pharmaceutical composition has been eliminated, reduced or minimized. The composition may include, or be administered in combination with, any other compound that will complement and enhance the effects of tocotrienols with respect to the treatment of androgenetic alopecia by administering tocotrienols, such as monoglycerides, lignans, isoprenoids, polyphenols, flavonoids, carotenoids, mono- and oligosaccharides, niacin, and bioactive peptides.

[0176] The invention is further described with reference to the following non-limiting examples.

[0177] Examples

[0178] Example 1: Effects of dietary tocotrienols on glucose tolerance, hypertension, myocardial fibrosis, and renal sodium excretion in rats

[0179] Sprague-Dawley rats were fed a high-fat diet (22%) for 8 weeks to induce obesity.

[0180] like Figure 2 It was shown that dietary tocotrienols (25 mg / kg / d) improved tolerance to a glucose load (2 g / kg, ip) in obese rats compared to animals not receiving tocotrienols (n=9, p<0.01; Mathai et al., 2011).

[0181] like Figure 3 As shown in Figure 2, dietary tocotrienols (50 mg / kg / d) gradually reversed established hypertension in obese rats compared with a placebo control group. Figure 4 As shown, in Figure 3The reduction in blood pressure observed in the study was accompanied by a reduction in myocardial fibrosis, as measured by left ventricular hydroxyproline content.

[0182] like Figure 5 As shown, myocardial fibrosis induced by isoproterenol was inhibited by tocotrienol supplementation. Since this model of myocardial fibrosis did not increase blood pressure, it was shown that the antifibrotic effect of tocotrienols was independent of the antihypertensive effect.

[0183] like Figure 6 It was shown that dietary supplementation with tocotrienols increased renal sodium excretion in rats fed a high-fat diet. This is important because obesity is often accompanied by sodium and water retention, which can lead to comorbidities such as increased blood volume and blood pressure and cardiovascular disease.

[0184] Example 2: Tocotrienol supplementation enhances motor performance in rats

[0185] Fifteen Sprague-Dawley rats were made obese by feeding a high-fat diet for 10 weeks and then divided into a control (n=7) and a tocotrienol-supplemented group (n=8). Figure 7 As shown, supplementation of the diet with a tocotrienol-rich fraction nearly doubled the ability of untrained rats to exercise on a treadmill.

[0186] In the first part of this experiment, an ascending test to exhaustion was performed to determine the maximum running speed of rats in the control and tocotrienol groups. Figure 7 A shows the mean peak speeds achieved by the control and tocotrienol-supplemented (50 mg / kg / d) groups during the exhaustion incremental test at week 8. This shows that the maximum speeds between the groups were similar. During the second treadmill run, individual rats ran at 65% of their maximum speed. Figure 7 B shows the average time taken to reach fatigue during the exhaustion time of the two groups at week 10. Data are shown as mean±SEM (n=7-8 / group). "*" indicates a statistically significant difference compared with the control group (P<0.05).

[0187] These results complement previous results showing that normal Wistar rats more than doubled their swimming exercise time when fed tocotrienols (Lee, SP, Mar, GY & Ng, LT 2009, "Effects of tocotrienol-rich fraction on exercise endurance capacity and oxidative stress in forced swimming rats", European Journal of Applied Physiology, 107, 587-595).

[0188] Example 3: Tocotrienol supplementation enhances exercise endurance in humans

[0189] Tocotrienols were formulated into the following sublingual dosage form: A 220 mg sublingual tablet formulation was prepared containing a 10 mg dose of tocotrienols consisting of the active ingredients delta-tocotrienol 9 mg and gamma-tocotrienol 1 mg, and the following excipients: dextrose, mannitol, marine gelatin, l-leucine, l-arginine, pregelatinized starch, PEG-200, propylene glycol, talc, silicon dioxide, magnesium stearate, carotenoids.

[0190] Four subjects were given two doses of 20 mg sublingual tocotrienol tablets in the morning and evening for a total of four weeks, followed by placebo in the same manner during an additional four-week period between and after tocotrienol treatment.

[0191] At the end of each four-week period, subjects performed an exhaustive incremental exercise test that consisted of cycling on a stationary bicycle with a stepwise increase in maximal effort (65%, 70%, 75%, 80%, 85%, adjusting each participant's maximum heart rate according to age) on a standard exercise bike. Subjects were equipped with a heart rate monitor and Borg ratings of physical exertion and time to exhaustion (range of 11-20) were obtained every 5 min during the cycling exercise.

[0192] Tocotrienol supplementation increased mean cycling time by approximately 40% (mean 133 minutes) compared to placebo treatment (mean 92 minutes). Figure 8 Subjective measures of exhaustion were also reached more slowly during increased exercise intensity, and total calculated distance traveled was greater during tocotrienol treatment (51 km) compared to placebo (32 km).

[0193] These results are in contrast to the only publication on human exercise, which showed no increase in exercise time to exhaustion on a treadmill in healthy young men supplemented with either placebo or 50 mg daily oral doses of palm-based tocotrienols for 6 weeks, despite increases in plasma antioxidant status (Keong CC, Singh HJ & Singh R. "Effects of palm vitamin E supplementation on exercise-induced oxidative stress and endurance performance in the heat," Journal of Sports Science and Medicine 5, 629-639).

[0194] Together, these two studies demonstrate that the sublingual route is important for translating the beneficial effects observed in rats via oral administration to effective results in humans.

[0195] Example 4: Tocotrienol supplementation reduces post-exercise soreness and delayed onset muscle soreness (DOMS):

[0196] Subjects administered the sublingual formulation of tocotrienols described in Example 2 were evaluated for post-exercise soreness and DOMS.

[0197] DOMS was induced by slow eccentric contraction of the forearm in four subjects. The subjects sat down and placed their forearms and elbows on the table. They were asked to lower a dumbbell of 10-12 kg (depending on the ability to control the weight) from a vertical arm position to the table, taking 3 seconds. This was repeated for 3 sets of 12 repetitions each with a one-minute rest period between each set. At the end of the 3-week period of supplementation with sublingual tocotrienols (40 mg / d) or placebo sublingual tablets, each subject underwent this test in a blind and random order.

[0198] Pain levels were assessed at 24 and 48 h after exercise by two methods: a visual analog scale on a 10-cm line on which subjects were asked to rate their pain level between 1 - no pain and 10 - extreme pain; in addition, subjects were asked to match their sensations with descriptive phrases: no pain, dull pain, mild pain, more mild pain, moderate pain, severe pain, extreme pain. Fig.10Shown are the visual analogue scales for 4 subjects who experienced visually no pain after tocotrienol supplementation (0.25 at 24h and 0 at 48h), compared to a rating of mild to more mild pain after placebo supplementation (2.25 at 24h and 48h).

[0199] In addition, subjects also volunteered pain ratings after exercise to exhaustion and after occasional exercise during the tocotrienol and placebo treatment periods. Subjects rated their pain levels after exercise to exhaustion, were consistently tired after both supplements after exercise to exhaustion, but experienced descriptions of "muscle stiffness," "pain for the next few days," "tired and can no longer walk," "adductor muscle stiffness and pain" only when taking the placebo supplement.

[0200] During casual exercise, subjects reported less pain and recovered more quickly from exercise during tocotrienol supplementation compared to the placebo period.

[0201] Example 5: Increased presence of tocotrienol concentrate in plasma following sublingual delivery of tocotrienol in gel form.

[0202] A gel formulation for sublingual delivery of tocotrienols was prepared as follows: Carboxymethylcellulose 2% and polyethylene glycol (0.5%) were placed in water to form a gel, which was then mixed with DeltaGold (70% pure tocotrienols extract based on annatto) to produce a 20 mg / ml tocotrienol gel.

[0203] One test was conducted in 2 subjects who were given 40 mg of tocotrienols in the form of a gel sublingually in 2 ml. Blood samples were collected from a forearm venous catheter before administration and at 15, 60 and 180 minutes after administration.

[0204] like Fig. 9As shown, tocotrienols rapidly appeared in plasma, with the level of delta isomer being 15 μM at 15 minutes after administration, decreasing to 3.2 μM, 1.5 μM, and 0.4 μM at 60 minutes, 120 minutes, and 180 minutes after administration. The levels of delta and gamma tocotrienols in plasma maintained their 9:1 tablet composition ratio, indicating that both tocotrienols were equally well absorbed via the sublingual route. This is in contrast to the oral route, where there is a difference in the appearance of tocotrienols in plasma, in the following order: alpha-tocotrienol > gamma-tocotrienol > delta-tocotrienol (Fairus S. et al., "Alpha-tocotrienol is the most abundant tocotrienol isomer circulated in plasma and lipoproteins after postprandial tocotrienol-rich vitamin Esupplementation", Nutrition Journal 2012, 11:5).

[0205] Example 6: Increased presence of tocotrienol concentrate in plasma following sublingual delivery of tocotrienol in tablet form.

[0206] A 220 mg sublingual tablet formulation was prepared containing a dose of 10 mg tocotrienols consisting of the active ingredients 9 mg of delta-tocotrienol, 1 mg of gamma-tocotrienol; and the following excipients: dextrose, mannitol, marine gelatin, l-leucine, l-arginine, pregelatinized starch, PEG-200, propylene glycol, talc, silicon dioxide, magnesium stearate, carotenoids.

[0207] One test was performed in 2 subjects who dissolved 4 x 10 mg tablets sublingually. Blood samples were collected from a forearm venous catheter before and after dissolving these tablets (20 min); and then at hourly intervals for 5 hours.

[0208] like Fig. 9 As shown, at the 20-minute time point, delta and gamma-tocotrienol appeared in plasma at high levels (8.2 μM for delta-tocotrienol and 0.8 μM for gamma-tocotrienol) and decreased over the next 3 hours (3.8 and 0.4 μM at 1 h; 1.7 and 0.19 μM and 0.02 at 2 h; 0.15 and 0.07 μM at 3 h).

[0209] Although the appearance of gel tocotrienols in plasma appeared greater initially compared to tablets, this was an artifact of the timing of sampling, as shown by the achievement of equal plasma levels by both tablet and gel formulations at later time points. During the depletion ramp-up test, the tablets were effective in the parameters we measured of reducing DOMS and increasing cycling time.

[0210] An important difference between oral administration and the two sublingual formulations is that tocotrienols appear rapidly in plasma within minutes compared to about 2 hours for the oral route. In addition, the observed CMax levels of sublingual tocotrienols at a dose of 40 mg (although from a small number of participants) are greater than the CMax levels achieved by oral ingestion of a higher dose (200 mg of tocotrienols alone; Hay YK, et al.).

[0211] Example 7: Tocotrienol supplementation reduces inflammation

[0212] A 220 mg sublingual tablet formulation was prepared containing a dose of 10 mg tocotrienols consisting of the active ingredients 9 mg of delta-tocotrienol, 1 mg of gamma-tocotrienol; and the following excipients: dextrose, mannitol, marine gelatin, l-leucine, l-arginine, pregelatinized starch, PEG-200, propylene glycol, talc, silicon dioxide, magnesium stearate, carotenoids.

[0213] One subject was given two doses of a 20 mg sublingual tocotrienol tablet in the morning and evening for a total of four weeks. The subject reported a reduction in inflammation and relief of pain from oral abscesses.

[0214] The same subjects with benign prostatic hypertrophy reported increases in urine flow and volume during tocotrienol supplementation, which gradually returned to their previous lower levels after discontinuation of tocotrienol supplementation.

[0215] Those skilled in the art will recognize that changes and modifications will become apparent. All such changes and modifications that become apparent to those skilled in the art should be considered to fall within the spirit and scope of the invention as described.

Claims

1. Use of a pharmaceutical composition in the preparation of a medicament for improving exercise endurance and behavioral performance in animals, wherein: The medicament is formulated for transmucosal administration into the salivary mucosal environment of a subject, The pharmaceutical composition comprises at least one tocotrienol and one or more pharmaceutically acceptable excipients, wherein the at least one tocotrienol comprises γ-tocotrienol and δ-tocotrienol in a ratio of 1:9, and the one or more pharmaceutically acceptable excipients comprise polyethylene glycol; and the composition is a solid dosage form, wherein the composition is provided in a form suitable for transmucosal administration in a salivary mucosal environment of a human, and the transmucosal administration in a salivary mucosal environment is sublingual administration or buccal administration.

2. The use according to claim 1, wherein The composition includes a tocotrienol-rich vitamin E portion including at least one tocopherol, wherein the vitamin E portion has a tocopherol component lower than a tocotrienol component.

3. The use according to claim 2, wherein The tocotrienol-rich vitamin E portion contains less than 10% tocopherol.

4. The use according to claim 1, wherein The composition does not include tocopherol.

5. The use according to claim 1, wherein The composition further comprises a tocotrienol selected from the group consisting of α-tocotrienol, β-tocotrienol, and combinations thereof.

6. The use according to claim 1, wherein The composition is in the form of a tablet, film, wafer, gum, or powder.

7. The use according to claim 6, wherein The composition is in the form of tablets or powders.

8. The use according to claim 1, wherein The one or more pharmaceutically acceptable excipients are dextrose, mannitol, marine gelatin, L-leucine, L-arginine, pregelatin starch, PEG-200, propylene glycol, talc, silicon dioxide, magnesium stearate, and carotenoids.

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