Annatto Extract Compositions, Including Geranyl Geraniols And Methods Of Use

ABSTRACT

Annatto extract composition (AEC), including cis and trans geranyl geraniols (GG) and tocopherol-free C-5 unsubstituted tocotrienols (T3), increases the de novo synthesis of intermediate isoprenoid and distal protein products, including endogenous coenzyme Q10 (CoQ10), dolichols (DL) and all subsequent GG-prenylated and DL-glycosylated proteins, including GG-porphyrinated hemes. This intermediate and distal product replenishment by AEC reverses maladies of myotoxicity (of both drug and non-drug origins), including maladies that affect the muscle, kidney, eye, GI tract and skin, nerve, blood, and CoQ10-related syndromes of energetics and LDL protection. AEC anabolically increases the endogenous de novo CoQ10 synthesis via GG elongation/prenylation of side-chain and conversely CoQ10 catabolically increases the endogenous de novo GG synthesis via beta-oxidation of CoQ10. Also, such AEC decreases de novo synthesis and increases disposal of triglycerides (TG) in humans via PPAR activation and SREBP deactivation. Such drop in TG by AEC reverses maladies of insulin resistance (IR) and metabolic syndrome (MS), prediabetes, diabetes and diabetes-related cardiovascular diseases (CVD). GG activates PPAR and down regulates SREBP transcription factors. This AEC, containing GG, inhibits cancer growth whether or not GG involvement in protein prenylation is required.
 RELATED APPLICATIONS

This application is a Continuation-in-Part of U.S. Non-provisionalapplication Ser. No. 10/821,679 filed on Apr. 8, 2004 (pending), whichclaims priority upon U.S. provisional application Ser. No. 60/461,612filed on Apr. 8, 2003 and claims priority upon U.S. provisionalapplication Ser. No. 60/528,353 filed on Dec. 10, 2003, the contents ofwhich are all herein incorporated by this reference in their entireties.All publications, patents, patent applications, databases and otherreferences cited in this application, all related applicationsreferenced herein, and all references cited therein, are incorporated byreference in their entirety as if restated here in full and as if eachindividual publication, patent, patent application, database or otherreference were specifically and individually indicated to beincorporated by reference.
 OTHER REFERENCES

-   Adachi, J. and G. Ioannidis (2000). “Glucocorticoid-induced    osteoporosis.” Drug Development Research 49: 120-134.-   Amin, D., R. Rutledge, et al. (1997). “RPR 107393, a potent squalene    synthase inhibitor and orally effective cholesterol-lowering agent:    comparison with inhibitors of HMG-CoA reductase.” J Pharmacol Exp    Ther. 281(2): 746-752.-   Ayanian, J., C. Fuchs, et al. (1988). “Lovastatin and    rhabdomyolysis.” Ann Intern Med. 109: 682-683.-   Baker, S. and M. Tarnopolsky (2001). “Statin myopathies:    pathophysiologic and clinical perspectives.” Clin Invest Med. 24(5):    258-272.-   Ballantyne, C., A. Corsini, et al. (2003). “Risk for myopathy with    statin therapy in high-risk patients.” Arch. Intern. Med. 163(5):    553-564.-   Bentinger, M., J. Grunler, et al. (1998). “Phosphorylation of    farnesol in rat liver microsomes: properties of farnesol kinase and    farnesyl phosphate kinase.” Arch Biochem Biophys 353(2): 191-198.-   Bi, X., M. Baudry, et al. (2004). “Inhibition of geranylgeranylation    mediates the effects of 3-hydroxy-3-methylglutaryl (HMG)-CoA    reductase inhibitors on microglia.” J. Biol. Chem. 279(46):48238-48245.-   Bliznakov, E. (2002). “Lipid-lowering drugs (statins), cholesterol,    and coenzyme Q10. The Baycol case—a modern Pandora's box.” Biomed    Pharmacother. 56(1): 56-59.-   Cenedella, R. (1995). “Role of transcription, translation, and    protein turnover in controlling the distribution of    3-hydroxy-3-methylglutaryl coenzyme A reductase in the lens.” Invest    Opthalmol Vis Sci. 36(10): 2133-2141.-   Cenedella, R. (1997). “Posttranscriptional regulation of    3-hydroxy-3-methylglutaryl coenzyme A reductase in lens epithelial    cells by mevalonate-derived nonsterols.” Exp Eye Res. 65(1): 63-72.-   Chamberlain, L. (2001). “Inhibition of isoprenoid biosynthesis    causes insulin resistance in 3T3-Liadipocytes.” FEBS Letters 507(3):    357-361.-   Ciosek, C., D. Magnin, et al. (1993). “Lipophilic    1,1-bisphosphonates are potent squalene synthase inhibitors and    orally active cholesterol lowering agents in vivo.” J. Biol. Chem    268: 24832-24837.-   Colli, S., S. Eligini, et al. (1997). “Vastatins inhibit tissue    factor in cultured human macrophages. A novel mechanism of    protection against atherothrombosis.” Arterioscler Thromb Vasc Biol    17: 265-272.-   Craveiro, A. A., et al., (1989). “The Presence of Geranylgeraniol in    Bixa Orellana Linn.,” Quimica Nova, 12(3):297-298.-   Cruz, A. and B. Gruber (2002). “Statins and osteoporosis: Can these    lipid-lowering drugs also bolster bones?” Cleveland Clinic Journal    of Medicine 69(4): 277-288.-   Danesi, R., C. McLellan, et al. (1995). “Specific labeling of    isoprenylated proteins: application to study inhibitors of the    post-translational farnesylation and geranylgeranylation.” Biochem    Biophys Res Commun. 206(2): 637-643.-   DeFronzo, R. A. (1998). “Pathogenesis of Type 2 Diabetes: Metabolic    & Molecular Implications for Identifying Diabetes Genes.” Ann. Rev. Diabetes: 1-93.-   Flint, O., B. Masters, et al. (1997).  “HMG CoA reductase    inhibitor-induced myotoxicity: pravastatin and lovastatin inhibit    the geranylgeranylation of low-molecular-weight proteins in neonatal    rat muscle cell culture.” Toxicol Appl Pharmacol. 145(1): 99-110.-   Flint, O., B. Masters, et al. (1997). “Inhibition of cholesterol    synthesis by squalene synthase inhibitors does not induce    myotoxicity in vitro.” Toxicol Appl Pharmacol 145(1): 91-98.-   Folkers, K., A. Osterborg, et al. (1997). “Activities of vitamin Q10    in animal models and a serious deficiency in patients with cancer.”    Biochem Biophys Res Commun. 234(2): 296-299.-   Gavish, D., E. Leibovitz, et al. (2000). “Bezafibrate and    simvastatin combination therapy for diabetic dyslipidaemia: efficacy    and safety.” J. Intern. Med. 247: 563-569.-   Grünler, J., J. Ericsson, et al. (1994). “Branch-point reactions in    the biosynthesis of cholesterol, dolichol, ubiquinone and prenylated    proteins.” Biochim Biophys Acta. 1212(3): 259-77.-   Guillet-Deniau, I., et al. (2003). “Glucose induces de novo fatty    acid synthesis in rat skeletal muscle through a SREBP-1c dependent    pathway.” Diabetes 52(Suppl. 1): extended abstract. 1024P.-   Hamilton-Craig, I. (2001). “Statin-associated myopathy.” Med J Aust. 175(9): 486-489.-   Haslinger, B., M. Goedde, et al. (2002). “Simvastatin Increases    Fibrinolytic Activity in Human Peritoneal Mesothelial Cells    Independent of Cholesterol Lowering.” Kidney International 62(5):    1611-1619.-   Hevener, A., W. He, et al. (2003). “Muscle-specific Pparg deletion    causes insulin resistance.” Nat. Med. 9(12): 1491-1497.-   Holdaas, H., A. Jardine, et al. (2001). “Effect of fluvastatin on    acute renal allograft rejection: a randomized multicenter trial.”    Kidney Int. 60(5): 1990-1997.-   Johnson, T. E., X. Zhang, et al. (2004). “Statins induce apoptosis    in rat and human myotube cultures by inhibiting protein    geranylgeranylation but not ubiquinone.” Tox. Appl. Pharma. 200:237-250.-   Johnston, S. and L. Kelland (2001). “Farnesyl transferase    inhibitors—a novel therapy for breast cancer.” Endocr Relat Cancer. 8(3): 227-235.-   Jondiko, I. J. O. and Pattenden, G., (1989). “Terpenoids and an    Apocarotenoid from Seeds of Bixa Orellana,” Phytochemistry,    28(11):3159-3162.-   Judy, W., T. Nguyen, et al. (2004). “Evidence for regression of PSA    and tumor size in patients with prostate cancer.” Submitted for    publication in Proc. Natl. Acad. Sci.-   Kotti, T. J., D. M. Ramirez et al. (2006). “Brain cholesterol    turnover required for geranylgeraniol production and learning in    mice.” Proc. Natl. Acad. Sci. USA 103(10):3869-74.-   Kraegen, E. (1998). “Physiologic manifestations of PPAR-gamma    activation: preclinical studies.” Clinical Courier 16(48): 5-7.-   Lankin, V., A. Tikhaze, et al. (2000).  “Intensification in vivo of    free radical oxidation of low density lipoproteins in plasma from    patients with myocardial ischemia treated by HMG-CoA-reductase    pravastatin and suppression of lipid peroxidation by ubiquinone    Q10.” Biull Eksp Biol Med. 129(2): 176-179.-   Lehmann, J., L. Moore, et al. (1995). “An antidiabetic    thiazolidinedione is a high affinity ligand for peroxisome    proliferator-activated receptor gamma (PPAR gamma).” J Biol Chem. 270(22): 12953-12956.-   Luckman, S., F. Coxon, et al. (1998). “Heterocycle-containing    bisphosphonates cause apoptosis and inhibit bone resorption by    preventing protein prenylation: evidence from structure-activity    relationships in J774 macrophages.” J Bone Miner Res. 13(11):    1668-1678.-   McCarty, M. (1999). “Can correction of sub-optimal coenzyme Q status    improve beta-cell function in type II diabetics?” Med Hypotheses. 52(2): 397-400.-   McGuire, T. and S. Sebti (1997). “Geranylgeraniol potentiates    lovastatin inhibition of oncogenic H-Ras processing and signaling    while preventing cytotoxicity.” Oncogene 14: 305-312.-   McLaughlin, T., F. Abbasi, et al. (2003). “Prediction of IR with    plasma TG or TG/HDL ratio.” Diabetes 52(Suppl. 1): A224 extended    abstract. 962P.-   Miller, P. (2001). “Bisphosphonates for the prevention and treatment    of corticosteroid-induced osteoporosis.” Osteoporos Int. Suppl 3:    S3-S10.-   Miquel, K., A. Pradines, et al. (1998). “Competitive Inhibition of    Choline Phosphotransferase by Geranylgeraniol and Farnesol Inhibits    Phosphatidylcholine Synthesis and Induces Apoptosis in Human Lung    Adenocarcinoma A549 Cells.” J. of Biol. Chem. 273(40): 26179-26186.-   Olbricht, C., C. Wanner, et al. (1997). “Accumulation of lovastatin,    but not pravastatin, in the blood of cyclosporine-treated kidney    graft patients after multiple doses.” Clin Pharmacol Ther. 62(3):    311-321.-   Omar, M., J. Wilson, et al. (2001). “Rhabdomyolysis and HMG-CoA    reductase inhibitors.” The Annals of Pharmacotherapy 35(9):    1096-1107.-   Ownby, S. and R. Hohl (2002). “Farnesol and geranylgeraniol:    prevention and reversion of lovastatin-induced effects in NIH3T3    cells.” Lipids 37(2): 185-92.-   Pearce, B., R. Parker, et al. (1992). “Hypocholesterolemic activity    of synthetic and natural tocotrienols.” J Med Chem. 35(20):    3595-3606.-   Peters, J., N. Hennuyer, et al. (1997). “Alterations in lipoprotein    metabolism in peroxisome proliferator-activated receptor    alpha-deficient mice.” J Biol Chem. 272(43): 27307-27312.-   Poels, P. and F. Gabreels (1993). “Rhabdomyolysis: a review of the    literature.” Clin. Neurol. Neurosurg. 95: 175-192.-   Polyerino, A. and S. Patterson (1997). “Selective activation of    caspases during apoptotic induction in HL-60 cells. Effects Of a    tetrapeptide inhibitor.” J Biol Chem. 272(11): 7013-7021.-   Qureshi, et al., (1995). “Response of Hypercholesterolemic Subjects    to Administration of Tocotrienols,” Lipids, 30(12).-   Raner, G., A. Muir, et al. (2002).  “Farnesol as an inhibitor and    substrate for rabbit liver microsomal P450 enzymes.” Biochem Biophys    Res Commun. 293(1): 1-6.-   Regazzi, M., I. Iacona, et al. (1993). “Altered disposition of    pravastatin following concomitant drug therapy with cyclosporin A in    transplant recipients.” Transplant Proc. 25: 2732-2734.-   Rogers, M. (2000). “Statins: lower lipids and better bones?” Nature    Medicine 6(1): 21-23.-   Saito, J., J. Davis, et al. (1995). “Users of low-dose    glucocorticoids have increased bone loss rates: a longitudinal    study.” Calcif Tissue Int. 57(2): 115-119.-   Schlienger, R., W. Haefeli, et al. (2001). “Risk of cataract in    patients treated with statins.” Arch Intern Med. 161(16): 2021-2026.-   Sever, N., B. Song, et al. (2003). “Insig-dependent ubiquitination    and degradation of mammalian 3-hydroxy-3-methylglutaryl-CoA    reductase stimulated by sterols and geranylgeraniol.” J. Biol. Chem. 278(52):52479-52490.-   Smith, P., R. Eydelloth, et al. (1991). “HMG-CoA reductase    inhibitor-induced myopathy in the rat: cyclosporine A interaction    and mechanism studies.” J Pharmacol Exp Ther. 257(3): 1225-1235.-   Smith, S. (1998). “The molecular pharmacology of PPAR-gamma.”    Clinical Courier 16(48): 3-4.-   Song, B., R. A. DeBose-Boyd, et al. (2006). “Insig-dependent    ubiquitination and degradation of 3-hydroxy-3-methylglutaryl    coenzyme A reductase stimulated by delta- and    gamma-tocotrienols.” J. Biol. Chem. 281(35):25054-25061.-   Spach, D., J. Bauwens, et al. (1991). “Rhabdomyolysis associated    with lovastatin and erythromycin use.” West J. Med. 154: 213-215.-   Staels, B., J. Dallongeville, et al. (1998). “Mechanism of action of    fibrates on lipid and lipoprotein metabolism.” Circulation 98(19):    2088-2093.-   Stark, W., M. Blaskovich, et al. (1998). “Inhibiting    geranylgeranylation blocks growth and promotes apoptosis in    pulmonary vascular smooth muscle cells.” Am J Physiol. 275: L55-L63.-   Takahashi, N., T. Kawada, et al. (2002). “Dual action of isoprenols    from herbal medicines on both PPARgamma and PPARalpha in 3T3-L1    adipocytes and HepG2 hepatocytes.” FEBS Lett. 514(2-3): 315-322.-   Talley, N., A. Weaver, et al. (1992). “Onset and disappearance of    gastrointestinal symptoms and functional gastrointestinal disorder.”    Am J Epidemiol. 136(2): 165-177.-   Thai, L., J. S. Rush, et al. (1999). “Farnesol is utilized for    isoprenoid biosynthesis in plant cells via farnesyl pyrophosphate    formed by successive monophosphorylation reactions.” Proc Natl Acad    Sci 96(23): 13080-13085.-   Theriault, A., et al., (1999). “Tocotrienol: A Review of Its    Therapeutic Potential,” Clinical Biochemistry, 32(5):309-319.-   Thibault, A., D. Samid, et al. (1996). “Phase I study of lovastatin,    an inhibitor of the mevalonate pathway, in patients with cancer.”    Clin Cancer Res. 2(3): 483-491.-   VERIS (1989). The role of Vitamin E in exercise, Vitamin E Research    & Information Service (November).-   VERIS (1990).  BACKGROUND OF THE INVENTION

1. Field of the Invention

The invention is on the compositions and uses of the extract from theannatto seed and such extract that is annatto oil or oleoresincontaining non-saponifiables, especially non-saponifiable terpenoids.
2. Description of the Related Art

Tocotrienols generally are classified as farsnesylated chromanols (FC)and mixed terpenoids (US 2004-0202740 A1, Tan). Tocopherol andtocotrienol are believed to have beneficial effects because they act asantioxidants. Tocotrienols, in particular, have been documented topossess hypocholesterolemic effects as well as an ability to reduceatherogenic apolipoprotein B and lipoprotein plasma levels. Further,tocotrienols are believed to be useful in the treatment ofcardiovascular disease and cancer (Theriault, A., et al., “Tocotrienol:A Review of its Therapeutic Potential,” Clinical Biochemistry,32:309-319 (July 1999); and “Tocotrienols: Biological and HealthEffects,” in Antioxidant Status, Diet, Nutrition, and Health, Papas, ed.(CRC Press), pp. 479-496 (1999)). Delta-tocotrienol andgamma-tocotrienol, in particular, have been identified as effectivesuppressants of cholesterol activity (Qureshi, et al., “Response ofHypercholesterolemic Subjects to Administration of Tocotrienols,”Lipids, 30(12) (1995)), and in inducing apoptosis of breast cancer cells(Yu, et al., “Induction of Apoptosis in Human Breast Cancer Cells byTocopherols and Tocotrienols,” Nutrition and Cancer, 33(1):26-32(1999)).
Tocols, which includes tocopherols and tocotrienols, have severalsources, including several vegetable oils, such as rice bran, soybean,sesame and palm oils. Tocotrienols have been discovered in the seeds ofBixa orellana Linn, otherwise known as the achiote tree (Jondiko, I. S.,et al., “Terpenoids and an Apocarotenoid from Seeds of Bixa Orellana,”Phytochemistry, 28(11):3159-3162 (1989)). However, each source oftocotrienols and tocopherols generally contains more than a single tocolhomolog. For example, palm oil and rice bran oil generally include bothtocotrienols and tocopherols. Further, alpha-tocopherol has beenreported to attenuate certain effects of tocotrienols, such as thecholesterol-suppressive activity of gamma-tocotrienol (Qureshi, et al.,supra.). In addition, because of their structural similarity,tocotrienols and tocopherols can be difficult to separate.
 Geranyl geraniol (GG) includes acyclic diterpene alcohols and geranylgeraniated terpenoids, and occurs naturally in linseed oil and cedrelatoona wood and tomato fruit. Geranyl geraniol also has been discoveredto exist in the seeds of Bixa orellana (Craveiro, et al., “The Presenceof Geranyl geraniol in Bixa Orellana Linn,” Quimica Nova, 12(3):297-298(1989)). Potential uses for geranyl geraniol include synthesis ofco-enzyme Q₁₀, vitamin K and tocotrienols. It is believed to inhibitesterification of retinol into inactive retinyl esters and, therefore,may be used to improve skin desquamation and epidermal differentiation(U.S. Pat. No. 5,756,109, issued to Burger, et al. on May 26, 1998).Geranyl geraniol has been employed in conjunction with HMG-CoA reductaseinhibitors in treatment of elevated blood cholesterol (WO 99/66929 byScolnick, published Dec. 29, 1999). Geranyl geraniol also is suspectedto be useful for treatment of human prostate cancer (U.S. Pat. No.5,602,184, issued to Myers, et al. on Feb. 11, 1997).
 Bixa orellana Linn, otherwise known as the achiote tree, is a member ofthe Bixaceae family and is native to tropical America. It is growncommercially in other parts of the world, generally within 20° of theequator or more preferably within 15° of the equator. The seeds of Bixaorellana Linn are the source of a reddish-orange colorant, known asannatto, that contains bixin and orelline, both of which are carotenoidpigments. The colorant is used commonly in foods, dyes and polishes.Typically, annatto is extracted from dehusked seeds in an aqueouscaustic solution. The colorant is precipitated from aqueous causticsolution by addition of a suitable acid, such as sulfuric acid. Theprecipitated colorant is removed by filtration. Filtercake ofprecipitated annatto colorant is dried and milled to form a commercialproduct. An oily phase generally is separated from an aqueous causticphase by centrifugation or by settling. Alternatively, the annattocolorant can be extracted from seeds in an organic solvent, such ashexane, acetone, or an alcohol. Miscella containing color and byproductoil are allowed to cool sufficiently to precipitate the annattocolorant. The precipitate is separated as bottoms from the organicsolvent. The oily phase from the caustic or organic extractionsfollowing separation of the annatto precipitate generally are discardedas byproducts.
The phrase “annatto extract” is imprecise and is used by other authorsto define a wide array of compositions. Typically, these productscontain all the parts of the annatto seed or at least contain the bixinsthat are used for colorants. It is important to distinguish annattoextracts that contain bixins from the disclosed annatto extracts by theapplicants that do not contain bixins or are essentially free of bixins.
 U.S. Pat. No. 6,350,453 by Tan discloses the method of obtaining thenatural annatto extract, the byproduct solution of Bixa orellana seedcomponents. The final product of the method is a composition containingthe natural ratio of isomers of the principal components.
The disclosed method in Tan('453) uses a “a vegetable oil” or “a ricebran oil” (Column 4, Lines 34-42) in one example to “reduce viscosity”(Line 39). This vegetable oil or rice bran oil was a cooking oil(triglyceride (TG)) which contained traces or none of any tocotrienols,tocopherols or geranyl geraniols.
The vegetable oil was used as a lubricant for the distillation ofannatto extract as it becomes more viscous with successive distillations(or “passes”). This is shown in Examples 3 & 4 in Tan('453) with the useof 10% rice bran oil.
Careful Examination of Example 4 in Tan('453), going over Passes 1 and2, after rice bran oil was added, then the Pass 2 distillation,tocotrienol was enriched. The vegetable oil was never distilled with thetocotrienol because the rice bran oil served as a lubricant (viscosityreducer) and not an additive or diluent to the tocotrienol. Thetocotrienol enrichment (about 15% to 26.3% in Pass 2, Example 4 or about19.6% to 33.6% in Pass 2, Example 3) shows that the vegetable oil wasleft behind as residue and did not distill with tocotrienol. Thedifference in molecular weights of TG and tocotrienol made thisprocessing possible. In Example 3, the rice bran oil ended in residue ofPass 2 (accounting 32% of the total mass).
 The byproduct solution of Bixa orellana seed components is obtainedafter removing the bixins to produce “yellow cake” and has greatlyreduced levels of bixins. The byproduct solution is then distilled toobtain a 290-390 Dalton MW fraction. The 290-390 Dalton MW fraction isthe fraction that contains geranyl geraniol. The disclosed geranylgeraniols in the application are obtained from the 290-390 Dalton MWfraction of the byproduct solution of Bixa orellana seed components.
Geranyl geraniols are 290 Daltons in molecular weight and Bixins are390-425 Daltons in molecular weight. These molecular weights are easy tocalculate from the structures of these chemicals. Additionally, theirchemical structures show that geranyl geraniols have 1 oxygen groups andbixins have 4 oxygen groups.
Although the molecular weights of bixins (390-425 Daltons) over lap themolecular weights of tocotrienol and tocopherol (350-450 Daltons), theirchemical structure [bixins have 4 oxygen groups; tocotrienols andtocopherols have 2 oxygen groups] inhibits bixins (i.e., they areheavier because they contain the 4 large oxygen groups) distillationfrom the byproduct solution of Bixa orellana seed components and remainin the residue material The higher the number of oxygen groups amolecule contains, the less likely it is to distill. Geranyl geraniolhas 1 oxygen group, tocotrienols and tocopherols have 2 oxygen groups,bixins have 4 oxygen groups; and triglycerides have 6 oxygen groups (MW500-1,000 Daltons). Therefore, geranyl geraniol distills before thetocotrienols and tocopherols, and bixins and triglycerides (being theheaviest/densest) remain in the residue.
It has been discovered that byproduct solutions of Bixa orellana seedcomponents contain tocotrienols, including delta- andgamma-tocotrienols, and geranyl geraniol. In particular, it has beendiscovered that tocotrienols and geranyl geraniol are present in thebyproduct oily phase of annatto colorant from annatto seeds and,especially, from whole dehusked annatto seeds.
 A “byproduct solution of Bixa orellana seed components” is definedherein as a solution derived from Bixa orellana seed components having aconcentration of annatto colorant significantly reduced from that ofBixa orellana seeds themselves. Other common terms for byproductsolution used for commercial products include: oil-soluble annatto coloror annatto oil. Generally, the concentration of annatto colorant, whichis defined as bixins and other carotenoids, chemically modified, alteredor esterified, in byproduct solution of Bixa orellana seed is less thanabout two percent, by weight, such as between about 0.05 weight percentand about 2.0 weight percent.
Annatto extract composition (AEC) typically contains cis and transisomers of geranyl geraniol (GG) and tocopherol-free tocotrienols (T3)that are essentially delta and gamma isomer forms. Geranyl geraniolsbelong to a class of terpenoid, more specifically, diterpene isoprenoidscontaining four isoprene units. The GG may be all in the trans isomerform (only one form possible), and/or contain one or more of cis isomerforms, both of the trans and cis forms are endogenous nutrients;however, they are not vitamins in the classical sense. Both cis andtrans GG become substrates for many branch-point reactions needed in thesyntheses of downstream isoprenoid and distal protein products.
 Geranyl geraniol (cis and trans) has a molecular weight of 290 Daltons,which is much smaller than the tocopherols and tocotrienols, and bixinsin the annatto seed. Vitamin E, including tocopherols and tocotrienols,are typically 390-430 Daltons in molecular weight or more broadly350-450 Daltons in molecular weight, which includes tocopherols andtocotrienols without any methylated groups in the lower range andtocopherols and tocotrienols with fully methylated groups in the higherrange. [Alpha-Tocopherol=430, Beta-Tocopherol=417, Gamma-Tocopherol=417,Delta-Tocopherol=403, Alpha-Tocotrienol=424, Beta-Tocotrienol=411,Gamma-Tocotrienol=410, and Delta-Tocotrienol=396.]

Many physiologic nutrients of small molecular weight are produced fromthe mevalonate pathway that generates the “isoprenoid pool” (IP)products. Geraniol (G), farnesol (F), and GG are the examples of IPproducts containing two, three, and four repeating units of five-carbonisoprenes, respectively. Tocotrienols belong to the class of vitamin Ethat includes tocopherols. It is known that T3's lower cholesterol andtreat hypercholesterolemia (Pearce, Parker et al. 1992; Song,DeBose-Boyd 2006). Unlike GG, T3's are not endogenous nutrients, but areproduced by plants and have a condensed farnesol tail in its structure.
 Farnesol constitutes the last committed step to cholesterol synthesis,but GG is not required for cholesterol synthesis (Flint, Masters et al.1997; Flint, Masters et al. 1997). GG constitutes the first uncommittedstep to cholesterol synthesis, and therefore, the first committed stepsin the synthesis of CoQ10, dolichol (DL), heme porphyrin, andGG-prenylated and DL-glycosylated proteins (Baker and Tarnopolsky 2001).Both cis and trans isomeric GGs are required for endogenous isoprenoidsubstrates for downstream branch-point products (Grünler, Ericsson etal. 1994). Trans-GG is the precursor to all-trans CoQ10 synthesis, whichis involved in mitochondrial respiration. Cis-GG is the precursor to DL,DL-glycosylated proteins, and certain GG-prenylated proteins. Dolicholand GG tend to concentrate in the brain and liver but GG is ubiquitouslyfound in many tissues (Grunler, Ericsson et al. 1994). Proteins producedby DL-glycosylation and GG-prenylation will be directed (e.g.,structures of protein fold, targets of where it will be delivered, andanchors of how it will be recognized). Deficiency in GG and/or DL leadsto improper localization of proteins, producing nonsense proteins andsignals. A major use of GG-prenylated protein is in the muscle tissues,and a major use of DL-glycosylated protein is in the nerve tissues.Synthesized proteins via isoprenoid GG and DL are described.
 The HMG CoA reductase (HMGR) catalyzes the rate-limiting steps in thelengthy hepatic cholesterol synthesis. The inhibition of HMGR is thetarget for statin targetment of hypercholesterolemia. However, statinsinhibit mevalonate (e.g., one isoprene) at the onset of the formation ofthe first isoprene, and therefore inhibits all subsequent IP products,including GG (FIG. 1). It is this depletion and deprivation of GG thatcan produce secondary, but clinically significant, side effects ofDL-starved cranial nerve damage and defects typified by neurologicaldysfunctions (e.g., taste alteration/loss, lack coordination, facialparesis, memory loss, vertigo, peripheral neuropathy, and peripheralnerve palsy). Geranyl geraniol salvages GG-prenylated proteins in braincells (That, S. Rush et al. 1999). Brain cells utilize free GG (not inthe activated GG-diphosphate form: GGPP) to restore the IP pool andincorporate it into the protein biosynthesis system. Thus, GG isphysiologically and pharmacologically significant in the central nervoussystem (CNS) (Kotti, T. J., D. M. Ramirez, et al 2006; Sever, N. B.Song, et al 2003; Bi, X., M. Baudry, et al 2004). For example, whenisoprenoid products are depleted by statin and bisphosphonatemedication, GG replenishes GG-prenylated and DL-glycosylated proteins.Drug side effects are many and they include GG-deprived induction ofmyotoxicities (e.g., musculoskeletal disorders, muscle cramps/pain,myalgia, myopathy, rhabdomyolysis, and myonecrosis), exo- andendothelial dysfunctions (e.g., upper GI maladies—esophagitis,gastritis/stomatitis, stomach/duodenal ulcer and lower GImaladies—constipation, dyspepsia, gastric dysmotility, abdominal pain)(Watts, Freedholm et al. 1999). GI tract (i.e., esophageal, gastric,duodenal) lesions include perforations, ulcers, bleeds and hemorrhages,maladies all of which come from GG-deprived protein synthesis of themucosae. Other GG-deprived dysfunctions include ocular maladies (e.g.,cataract/lens opacity, dry eyes, corneal abrasion, ophthalmoplegia),anemia, CoQ10, DL and its associated DL-starved maladies, describedabove. Again, eye problems such as lens opacity and dry eyes can betraced to the deprivation of GG. These side effects include secondaryCoQ10-deprived maladies (e.g., mitochondrial dysfunction,ATP/respiration, LDL protection, tiredness/malaise, fibromyalgia,chronic fatigue syndrome, and congestive heart failure). The schematicoutline of this invention for GG-deprived maladies is shown in FIG. 2.
 Drug-Induced Myopathies Via GG Inhibition

IP product depletion from treatment with statins is serious side effect,so alternatives to statins are proposed for treatment ofhypercholesterolemia. Squalene synthase catalyses the first committedstep in cholesterol biosynthesis via two F groups head-to-head (FIG. 1).To avoid such global IP depletion, and particularly GG depletion,squalene synthase inhibitors (SSI) target distal isoprenoid squaleneinhibition to treat hypercholesterolemia (Ciosek, Magnin et al. 1993;Amin, Rutledge et al. 1997). A unique advantage of SSI, as opposed tostatins, is that they do not deplete IP immediate and distal products,such as GG, CoQ10, and DL. Such new drug targets only underscore theunique role of GG and the serious implication of its depletion. However,widespread successful use of statins, and their ever growing expandeduses, emphasizes the importance of the invention for adjunctive therapyto circumvent isoprenoid depletion in general, and GG depletion inparticular (Johnson, T. E., X. Zhang, et al 2004).
Isoprenoid pool deprivation and myopathies are common with widespreaduse of statin drugs for the treatment of hypercholesterolemia, fibratedrugs for the treatment of hypertriglyceridemia, and bisphosphonatedrugs for the treatment of osteoporosis. Such widespread use of statinsis now extended further because of other non-cholesterol approved uses,other cardiovascular indications/uses, as well as, otherstatin-in-tandem combination uses. A clinically meaningful adverse eventof GG inhibition is a global loss of protein, with consequentmyotoxicity. Therefore, AEC is particularly useful in the adjunctiverelief to IP deprivation, such as, but not limited to statin, fibrate,and bisphosphonate users.
Non-Drug-Induced Myopathies Via GG Inhibition

Isoprenoid pool deprivation may also occur in the elderly and those withAIDS-HIV where wasting occurs due to protein deficit (Poels and Gabreels1993; Hamilton-Craig 2001).
 CoQ10

CoQ10 is transported in the vascular system via LDL particles. Statinswork to inhibit de novo cholesterol synthesis, which also simultaneouslyinhibit de novo CoQ10 and DL synthesis (Bliznakov 2002). Statins alsowork to increase the hepatic LDL receptors, hence reducing LDL particlesin vascular circulation. Consequently, patients on statins will see adrop in LDL with a corresponding drop in CoQ10 (Watts, Castelluccio etal. 1993).
GG is the first committed step for numerous downstream distal products,including CoQ10 (FIGS. 1 & 4). The GG molecule (MW=290) containing 4isoprene units anabolizes to CoQ10 molecule (MW=863) containing 10isoprene units. Conceptually, a minimum of 2 moles of GG is required toanabolize 1 mole of CoQ10 and conversely 1 mole of CoQ10 is required tocatabolize to 2 moles of GG. This is illustrated by way of the molarconversion example as follows: A 100 mg of GG (100/290=0.345 mmole) cananabolize to 150 mg of CoQ10 (0.345/2×863).
Hypercholesterolemia

Statin intensifies in vivo LDL oxidation in patients with myocardialischemia while CoQ10 supplementation suppresses lipid oxidation (Lankin,Tikhaze et al. 2000). Further, animal cells contain about 10-fold moreCoQ10 than vitamin E, and the cell preferentially utilizes CoQ10 as anantioxidant.
This invention shows AEC supplementation prevents statin toxicities,increases CoQ10, and the endogenous CoQ10 preferentially protects theLDL, lowers cholesterol and improves endothelial functions all at thesame time. For patients on statins, endogenous CoQ10 levels typicallydrop about 30-40%. Clinically significant adverse effects occur whenCoQ10 levels fall below 0.5 ug/mL. AEC also help diabetics on statins byenhancing CoQ10 status which improves beta-cell function in Type 2diabetes (McCarty 1999).
 It is implicit to current discussions that GG is readily bioavailable tocells and tissues. In addition, GG is not cytotoxic as it does not causecell rounding, a known cellular indicator of myotoxicity (McGuire andSebti 1997; Ownby and Hohl 2002). In fact, GG prevents and reverses cellrounding caused by statins and bisphosphonates. However, a similar IPproduct, farnesol, does not have either of these GG benefits. Therefore,the use of AEC takes advantage of the bioavailability and safety of GGto tissues.
Statin inhibits the insulin-responsive glucose transporter (Glut 4), andthat such inhibition of IP biosynthesis cause IR in adipocytes(Chamberlain 2001). Glut 4 is a membrane protein that requiresGG-prenylation. Therefore, the use of statins and bisphosphonates wouldinhibit the GG-prenylated biosynthesis of Glut 4, and thereby causinginsulin resistance (IR) in adipocytes.
Cancer

A strategic way to inhibit cancer is to employ a farnesyl transferaseinhibitor (FTI), since Ras cancer requires farnesyl-prenylation of itsprotein for survival. These FTIs are known to have toxic effects tocancer patient including GI toxicity, peripheral neuropathy & nerveconduction abnormality, and fatigue (Johnston and Kelland 2001).Surprisingly, all of these toxic effects may be ascribed to GGdeficiency. GI toxicity is due in part to GG-associated prenylation ofprotein on the GI lining. Neuropathy and nerve defects are often relatedto DL-depleted glycosylation. Fatigue is often of unknown etiologies,commonly associated with chronic fatigue syndromes. They are ascribed toa deficiency in CoQ10, derived endogenously from the GG substrate.
Statin drugs have also been used in cancer treatment. A typical dosageof statins for cancer is 10 times their requirements for cholesterolreduction (Wong, Dimitroulakos et al. 2002). This can lead to seriousmyotoxicities including myopathy and rhabdomyolysis. GG is not toxic tountransformed cells or to normal cells (Stark, Blaskovich et al. 1998;Ownby and Hohl 2002; Wong, Dimitroulakos et al. 2002).
 Cancer patients often have low blood levels of CoQ10. CoQ10 has beenused as treatment in patients with breast and prostate cancers (Folkers,Osterborg et al. 1997; Judy, Nguyen et al. 2004). The prostate specificantigen (PSA) and prostate mass of prostate cancer patients after oneyear of CoQ10 supplementation decreased 71% and 47%, respectively.However, the mechanism of such effect is not yet known. Prostate cancerpatients taking up to 600 mg/day CoQ10 is equivalent to taking 400mg/day supplement of GG (Judy, Nguyen et al. 2004) according to earlieranalysis (see CoQ10 section).
CoQ10 reduces the severity but not the incidence of musculoskeletaltoxicities and patient complaints (Thibault, Samid et al. 1996; Wong,Dimitroulakos et al. 2002). Supplementation of mevalonate, a directprecursor to GG but not CoQ10, is shown to ameliorate myopathy,suggesting that the toxic effects are not due to CoQ10 deficiency(Smith, Eydelloth et al. 1991). These studies lend corroborative supportto the above claim that CoQ10 catabolizes to GG, at least in parts,which in turn is responsible for partial reversal of myopathy. It mayalso be understood that it is GG, not CoQ10 per se, reverses myopathy.
While many biological processes are anabolic in nature, catabolicprocesses are also well known. One such isoprenoid catabolism is theconversion of cholesterol to Vitamin D, steroid hormones, and bile acids(FIG. 2). Such a strategy of cancer treatment is unique, as both CoQ10and GG are endogenous nutrients, while the majority of cancer drugs arexenobiotic.
 There are numerous strategies that disclose the use of GG for cancertreatment, which directly or indirectly involve GG protein prenylation(McGuire and Sebti 1997; Ownby and Hohl 2002). However, its apoptosismechanism remains largely unknown. Two hypotheses come closest toexplaining the mechanism as a “common effector” or a “coordinatedregulator” of apoptosis by GG. GG results in a rapid en masse inductionof apoptosis via activation of caspase-3 and possibly caspase-2(Polyerino and Patterson 1997). GG very quickly induces phosphatidylcholine biosynthesis inhibition at the level of cholinephosphotransferase, the last step of CDP-choline known as the Kennedypathway (Miquel, Pradines et al. 1998). Surprisingly, neither of the twoapoptosis hypotheses require GG prenylation nor involve proteinsynthesis for apoptosis. GG appears to be the common denominator and avery potent compound to induce apoptosis en masse. It should be notedthat GGPP is not stable and is unlikely to penetrate cell membranesunaided, but the natural isoprenol GG is bioavailable, and taken up bycells through an active transport system, and/or dephosphorylatedsequentially by kinases (Danesi, McLellan et al. 1995; Bentinger,Grunler et al. 1998).
Renal Insufficiency

Renal insufficiency affects about 20 million Americans. The continuousirritation of the peritoneum in peritoneal dialysis patients can resultin local peritoneal fibrinolytic activities as measured by fibrinolyticenzyme tissue-type plasminogen activator (t-PA) and plasminogenactivator inhibitor-1 (PAI-1). Statins increase the t-PA and decreasethe PAI-1 and may cause defects in the actin cytoskeleton (Haslinger,Goedde et al. 2002), which may irritate and thin the peritoneal lining.It is noted that the negative effects of statins can be prevented orreversed by the use of GG (Colli, Eligini et al. 1997; Haslinger, Goeddeet al. 2002). Since many statins including cerivastatin, pravastatin,lovastatin, and simvastatin are filtered in part through the kidneys andexcreted as urine, these drugs can exasperate the problems of renalinsufficient patients.
 Organ Transplants

Annually there are approximately 2,000 heart and 14,000 kidneytransplants performed in the US. Patients with kidney and hearttransplants are normally given cyclosporine to suppress the immuneresponse to organ rejection. The most common side effects ofcyclosporine are kidney dysfunction and failure, as measured by elevatedblood creatinine and uric acid. These side effects may be caused bydecreased efficiency in the glomerular filtration rate (GFR), indicatingrenal insufficiency. Since most graft patients have elevated lipidlevels that can lead to coronary artery disease, statins are oftenprescribed along with cyclosporine. For these patients, the risks ofmyopathy and/or rhabdomyolysis are substantially higher (ca 15-80%).Despite the dangers of myotoxicities of this combo therapy, their usageis justified based on benefit-to-risk assessment provided that thestatin doses are on the lower end, only one statin is allowed, and nofibrates (Ballantyne, Corsini et al. 2003).
Myotoxicities

Myotoxicity includes all forms and stages of muscle damage including,but not limited to, myalgia, myopathy, and rhabdomyolysis. Myopathy isalso associated with generalized myalgia and recurrence of fatigue orweakness (creatine kinase level, CK>10 times the normal value).Rhabdomyolysis is characterized by global skeletal muscle fiberbreakdown. Organ damage, typically renal insufficiency or acute renalfailure, accompanies rhabdomyolysis when CK>100 times the normal value.
Myopathy and rhabdomyolysis may also have non-drug origins. Among thecommon causes that are not drug-induced are traumas (e.g. surgery),infections (e.g. viral, bacterial, and fungal), exercise exertion,alcohol abuse, and other inherited, environmental, or metabolic causes(Poels and Gabreels 1993; Hamilton-Craig 2001). Therefore, myotoxicityof both drug-induced and non-drug-induced origins are widespread asevidenced by the mild form, myalgia, to intermediate form, myopathy, toseverest form, rhabdomyolysis.
 There are many known causal mechanisms for drug-induced myopathiesincluding inhibitions of cytochrome 3A4, HMGR, GG, and P-glycoproteins.Statins and bisphosphonates are particularly effective inhibitors ofHMGR and GG. These two classes of drugs have remarkably overlappingmodes of action. For example, statins, known for its cholesterolreduction via HMGR inhibition, reduce osteoporosis (Rogers 2000; Cruzand Gruber 2002). Conversely, bisphosphonates, known for bonestrengthening via GG inhibition, reduce cholesterol (Ciosek, Magnin etal. 1993). Surprisingly, both statins and bisphosphonates inhibit cancervia FT inhibition (Luckman, Coxon et al. 1998; Wong, Dimitroulakos etal. 2002).
Most drugs are extensively biotransformed by the metallo-protein enzymecytochrome P450 (CYP) system, with the majority of them processed by CYP3A4, including statins. These processed drugs are removed from the bodythrough biliary and renal excretions in a safe manner. When enzymaticprocessing by CYP 3A4 is depressed, drug concentration (e.g. statin)becomes elevated in the blood. Such elevation can occur during statinmonotherapy or combo-therapy with erythromycin (where blood statinconcentration is known to increase by 3-8 folds) (Ayanian, Fuchs et al.1988; Spach, Bauwens et al. 1991) or with cyclosporine (where bloodstatin concentration is known to increase by 6-23 folds) (Regazzi,Iacona et al. 1993; Olbricht, Wanner et al. 1997; Holdaas, Jardine etal. 2001). Similar interactions can occur with other drug classes suchas warfarin, antifungals/antibiotics, and niacin. The resultant statinelevation in the vascular system can cause serious GG depletion, leadingto myopathy and rhabdomyolysis. It is important to note that GG does notinhibit any of the cytochrome P450 enzymes for which CYP3A4 is a part of(Raner, Muir et al. 2002).
 Fibrates are effective in lowering triglyceride and hence areparticularly useful for prediabetics and Type II diabetics; however,they tend to have a high toxic side effect of myopathy. For prediabeticand diabetic patients, benefits may outweigh the risk in combo therapywith statins to treat mixed lipidemia, common in this patient group.However, the incidence of myopathy may increase by 10-folds in diabeticsas compared to the general population when on combo therapy (i.e.myopathy increased from 0.12% to 1.35%) (Gavish, Leibovitz et al. 2000;Omar, Wilson et al. 2001).
Even in monotherapy, fibrates cause myopathy 5.5 times greater thanstatins, posing an independent risk for myopathy. Fibrates are excretedthrough the kidneys, which can cause serious problems even in peoplewith mild renal impairment.
Insulin Resistance

Insulin resistance (IR) is associated with increased risk ofcardiovascular disease (CVD), Type 2 diabetes mellitus (T2DM),hypertension, polycystic ovarian syndrome (PCOS) and alcohol-unrelatedfatty liver disease. However, plasma insulin measurement is notstandardized across clinical laboratories, and therefore is anunreliable marker. Therefore, a surrogate marker was developed forinsulin resistance, where the IR criteria are TG/HDL≦3.5 and/or TG≦140mg/dL (McLaughlin, Abbasi et al. 2003).
GG activates mixed PPARs, both PPARγ at the adipocytes and PPARα at thehepatocytes (Takahashi, Kawada et al. 2002). PPARγ activation in adiposetissues decreases IR (Lehmann, Moore et al. 1995; Willson, Lambert etal. 2001) and PPARα activation in the liver lowers blood lipids (Peters,Hennuyer et al. 1997; Staels, Dallongeville et al. 1998). Furthermore,statin down regulates glucose transporter 4 (Glut 4) expression andthereby suppresses the glucose uptake into cells with consequent IR(Chamberlain 2001). Therefore, IP products that are decimated by statininhibition may inhibit the GG-prenylated protein synthesis of Glut 4.
 Peroxisomal Proliferator Activated Receptors

Peroxisomal proliferator activated receptors (PPAR) are members of thenuclear receptor transcription factors. The metabolic consequences ofPPARγ activation have been researched mostly on adipose tissue where itis largely expressed (Kraegen 1998; Smith 1998), as well as, on muscletissue (Hevener, He et al. 2003). The metabolic effects of known PPARactivator thiazolidinedones (TZD) are, a) reduces hyperglycemia andhyperinsulinemia, b) lowers FFA and TG levels, c) enhances IS and lowersIR states, and d) requires insulin for glucose-lowering action. NumerousPPARγ activator functions are similar to PPARα activator functions. ThisPPARα has been actively researched on liver tissue, especially withregards to lipid use (e.g., uptake and beta-oxidation). Even though theaction sites of PPARγ (mainly in adipose) and PPARα (mainly in liver)are different, their activations have many overlapping outcomes.Typically TZD and fibrates affect the activation of PPARγ and PPARα,respectively.
Sterol Regulatory Element Binding Protein-1

Sterol regulatory element binding protein-1 (SREBP-1) is a transcriptionfactor that responds to nutritional status and regulates metabolic geneexpression in various organs, including liver, adipose and muscle. Ithas been shown that insulin and glucose induces de novo fatty acidsynthesis leading to a rapid increase in lipogenic flux in skeletalmuscle. This lipid accumulation is associated with muscle IR in obesityand T2DM, and is stimulated/mediated via the SREBP-1 expression(Guillet-Deniau 2003). As discussed earlier, IR is tightly associatedwith increased lipids (McLaughlin, Abbasi et al. 2003) and increasedinsulin or hyperinsulinemia (HI) (DeFronzo 1998). Additionally, theSREBP-1 expression in part controls FFA/TG synthesis, and PPARexpression in part controls FFA/TG uptake and catabolism (Song, B., R.A. DeBose-Boyd 2006).
 Other Aspects of GG Deficiencies and Uses:

The upper GI track (esophagus, stomach, and duodenum) is particularlysensitive to perforations, ulcers, and bleeds. Collective adverse events(AE) include, but not limited to, abdominal pain, dyspepsia, esophagealerosion, esophagitis, reflux esophagitis, and the likes in the duodenum.Repairs to the GI track are done by cellular replication and takeapproximately 2 weeks in esophagus. Repairs by mucosal migration takeapproximately 2 days in the duodenum and 2 hours in the stomach.Therefore opportunistic AE is most likely to occur in the esophagusfollowed by duodenum and least likely in the stomach. Not surprisingly,drug-induced upper GI AE are common, especially in the esophagus. Thesedrugs include emepronium bromide, doxycycline, tetracycline antibiotics,iron supplements, quinidine, non-steroid anti-inflammatory drugs(NSAIDs), alprenolol, captopril, theophylline, zidovudine, andbisphosphonates. Studies show 20-30% of patients develop upper GI AEwithin the first year of bisphosphonate therapy (Talley, Weaver et al.1992). The mechanism of upper GI ulcer-related events is due to the GI'sinability to prenylate protein needed for cellular replication (a muchslower process than mucosal migration) caused by drug-induced depletionof GG and localized esophagitis caused by pills slipping through theesophagus (Watts, Freedholm et al. 1999)

Asymptomatic endoscopic abnormalities (e.g. hemorrhages, erosions, andulcers) are surprisingly high (15%) in normal post menopausal women(Watts, Freedholm et al. 1999).
 Steroids are widely used and the most common among them is prednisone.Corticosteroids are used for many inflammatory diseases including butnot limited to arthritis, connective tissue disease, asthma, and inheart transplant patients. These corticosteroids have several sideeffects including rapid loss of bone mass in the first year of use, ashigh as 15% of patients develop vertebral fractures (Adachi andIoannidis 2000), loss of bone mineral density even at very low doses,e.g. prednisone at 5 mg/day (Saito, Davis et al. 1995), and a high rateof steroid-induced osteoporosis, higher than osteoporosis in postmenopausal women (Miller 2001). To prevent and reversecorticosteroid-induced osteoporosis, bisphosphonates has become the bestdrug candidate.
The role of Vitamin E in exercise is well known. Muscle damage can occurduring exhaustive exercise, even in highly trained athletes.Furthermore, since the body's Vitamin E consumption increases with theamount of exercise, high amounts of Vitamin E are needed for endurancetraining and for membrane lipid oxidation protection during strenuousexercise (VERIS 1989).
Statins and bisphosphonates can increase the risk of adverse ocular sideeffects including cataracts (Schlienger, Haefeli et al. 2001). Statinsincrease the mRNA and the protein mass of HMGR, which translates to anover expression of cholesterol biosynthesis in intact lens (Cenedella1995; Cenedella 1997). It is suggested that IP products might preventlens opaqueness, cataract, and lens cholesterol deposition. Cataractremoval remains the most common surgery in the US (more than halfmillion per year). The occurrence of cataracts approaches 50% for those75 years or older. The protective use of Vitamin E against cataractdevelopment is well recognized (VERIS 1990). Vitamin E tocotrienols andtocopherols are both powerful antioxidants. However, only tocotrienols,especially delta- and gamma-tocotrienols have been shown to downregulate the mRNA and reduce the protein mass of HMGR.
 DEFINITIONS

Annatto extract—A source of material known as a byproduct solution ofBixa orellana seed components, which is obtained as an oily oleoresinousmaterial after the bulk of annatto color, is largely removed from eitherthe aqueous extract or solvent extract of annatto seeds. Further, thisbyproduct contains a tocotrienol component and a geranyl geraniolcomponent and can be used as a source for the recovery of a tocotrienolcomponent and a geranylgeraniol component. The annatto extract contains≦2% carotenoids, especially bixins, and mostly (30-40%) as lightvolatiles of MW<250, geranyl geraniols (20-40%) and tocotrienols(10-20%), and other heavy condensates (5-10%).
Annatto Extract Oil—The oily product from the annatto seed extractcontaining the tocotrienols and tocopherols, collectively tocochromanols(Vitamin E), which are in the 350-450 Dalton MW fraction of the extract.It is denuded or essentially free of carotenoids (bixins), lightvolatiles, and heavy condensates. Saponifiable vegetable cooking oils,commonly known as triglycerides, which have a range of molecular weightsof 500-1000 Daltons, and are essentially free or free of tocotrienolsand tocopherols (typically <0.05% Vitamin E) are not included withinthis definition.
Appropriate Spectrum of Tocols—Mixtures of annatto tocotrienols withother plant extracts to achieve efficacy of the newly constituted tocolsor tocochromanols composition. Annatto tocotrienols satisfy thisdefinition by having the highest amount of C5 unsubstituted tocotrienolsand the lowest amount of tocopherols, especially alpha-T1.
Chemotactic Bioactive Materials—Biochemical molecules involved in anyoxidative/inflammatory process that leads to loss of arterialvasculature.
Corn Oil—The oily product from the corn extract containing thetocotrienols and tocopherols, collectively tocochromanols (Vitamin E),which are in the 350-450 Dalton MW fraction of the extract. Saponifiablevegetable cooking oils, commonly known as triglycerides, which have arange of molecular weights of 500-1000 Daltons, and are essentially freeor free of tocotrienols and tocopherols (typically <0.05% Vitamin E) arenot included within this definition.
 Cottonseed Oil—The oily product from the cottonseed extract containingthe tocotrienols and tocopherols, collectively tocochromanols (VitaminE), which are in the 350-450 Dalton MW fraction of the extract.Saponifiable vegetable cooking oils, commonly known as triglycerides,which have a range of molecular weights of 500-1000 Daltons, and areessentially free or free of tocotrienols and tocopherols (typically<0.05% Vitamin E) are not included within this definition.
Cranberry Seed Oil—The oily product from the cranberry seed extractcontaining the tocotrienols and tocopherols, collectively tocochromanols(Vitamin E), which are in the 350-450 Dalton MW fraction of the extract.Saponifiable vegetable cooking oils, commonly known as triglycerides,which have a range of molecular weights of 500-1000 Daltons, and areessentially free or free of tocotrienols and tocopherols (typically<0.05% Vitamin E) are not included within this definition.
Essentially Free of Bixins—A composition that contains less than 0.1% byweight of bixins.
Essentially Free Of Tocotrienols—A composition that contains less than0.1% by weight of Tocotrienols.
Ingestible—descriptive of a manner to administer compressed tablets,softgel gelatin, hard gel two-piece gelatin, beads, granules, and/orliquid coats.
Litchi Seed Oil—The oily product from the litchi seed extract containingthe tocotrienols and tocopherols, collectively tocochromanols (VitaminE), which are in the 350-450 Dalton MW fraction of the extract.Saponifiable vegetable cooking oils, commonly known as triglycerides,which have a range of molecular weights of 500-1000 Daltons, and areessentially free or free of tocotrienols and tocopherols (typically<0.05% Vitamin E) are not included within this definition.
Medicament—a substance used in therapy.
 MW (Molecular Weight) Fraction—Refers to the part (fraction) of asubstance (i.e., natural extract) that has chemicals of that molecularweight. A standard analytical tool in Biochemistry or Chemistry is theseparation of a substance into its various individual chemicals by theirmolecular weight. Typical methods include column chromatography, HPLC,and SDS-PAGE. Each of these analytical tools will separate a complexsubstance (i.e., natural extract) so the individual chemicals willtravel at different rates through the medium (e.g., silica orcoated/reversed phase silica, sepharose beads or polymerized gels). Inthe case of column chromatography or HPLC, the carrier solution iscollected (e.g., test tubes) as it comes off the column into“fractions”. There is a detector on the end of the column which detectsthe presence of material. The detector charts the “peaks” and thecorresponding fraction that contains this material. The molecularweights of these peaks can be calculated using standards with knownmolecular weights (e.g., Keyhole Limpet Hemocyanin) or the purecompounds with previously identified MWs.
Oat Bran Oil—The oily product from the oat bran extract containing thetocotrienols and tocopherols, collectively tocochromanols (Vitamin E),which are in the 350-450 Dalton MW fraction of the extract. Saponifiablevegetable cooking oils, commonly known as triglycerides, which have arange of molecular weights of 500-1000 Daltons, and are essentially freeor free of tocotrienols and tocopherols (typically <0.05% Vitamin E) arenot included within this definition.
Olive Oil—The oily product from the olive extract containing thetocotrienols and tocopherols, collectively tocochromanols (Vitamin E),which are in the 350-450 Dalton MW fraction of the extract. Saponifiablevegetable cooking oils, commonly known as triglycerides, which have arange of molecular weights of 500-1000 Daltons, and are essentially freeor free of tocotrienols and tocopherols (typically <0.05% Vitamin E) arenot included within this definition.
 Palm Oil—The oily product from the palm extract containing thetocotrienols and tocopherols, collectively tocochromanols (Vitamin E),which are in the 350-450 Dalton MW fraction of the extract. Saponifiablevegetable cooking oils, commonly known as triglycerides, which have arange of molecular weights of 500-1000 Daltons, and are essentially freeor free of tocotrienols and tocopherols (typically <0.05% Vitamin E) arenot included within this definition.
Purification of Annatto Extract—A process to obtain isomers oftocotrienol and geranylgeraniol where the ratios are different fromfound in nature. Purified annatto extract is denuded or essentially freeof carotenoids (bixins), light volatiles, and heavy condensates.
Purified Annatto Geranylgeraniol Composition—A composition purified froman annatto extract that contains one or more of the isomers ofgeranylgeraniol in a ratio that is different from the natural ratiofound in an annatto extract. The trans:cis ratio of geranylgeraniolsfound naturally annatto seeds is within the range of 20:1 to 6:1.However, by using specialized methods, a purified annattogeranylgeraniol composition can be obtained that has altered trans:cisratios of geranylgeraniols from found in the natural annatto seed. Apurified annatto geranylgeraniol composition can have only trans isomer,essentially only trans isomer, and trans:cis ratios from 100:1 to 1:5.
Purified Annatto Tocotrienol Composition—A composition purified from anannatto extract that contains one or more of the isomers of tocotrienolin a ratio that is different from the natural ratio found in an annattoextract.
Rice Bran Oil—The oily product from the rice bran extract containing thetocotrienols and tocopherols, collectively tocochromanols (Vitamin E),which are in the 350-450 Dalton MW fraction of the extract. Saponifiablevegetable cooking oils, commonly known as triglycerides, which have arange of molecular weights of 500-1000 Daltons, and are essentially freeor free of tocotrienols and tocopherols (typically <0.05% Vitamin E) arenot included within this definition.
