Decoding The Elusive Vitamin E: Why Tocotrienol Trumps Tocopherol By Jerry Tan, M.d.

INSIDER Webinar Series presents

Decoding the Elusive Vitamin E: Why Tocotrienol Trumps Tocopherol Sponsored by

To register for additional free Webinars, visit NaturalProductsInsider.com

Today’s Webinar 

Today’s webinar will be available at NaturalProductsInsider.com on Tuesday, Feb. 3



To access this webinar, go to NaturalProductsInsider.com, click on Webinars and select today’s webinar title to view the presentation.

Today’s Presenter Barrie Tan, Ph.D., earned his bachelor’s degree in chemistry and doctorate in analytical chemistry at the University of Otago, New Zealand, and later became a professor of chemistry and food science/nutrition at the University of Massachusetts, Amherst. His research expertise included lipid-soluble materials such as carotenoids, tocotrienols/tocopherols, CoQ10, omega-3s and cholesterol. He was the first to introduce tocotrienol’s benefits to our nutrition industry. He founded American River Nutrition Inc. in 1998 and developed the first ever tocopherol-free tocotrienol product derived from annatto beans. Today, the focus of his research is on phytonutrients that have an impact on chronic, degenerative and cancer diseases.

© 1/09

Decoding the Elusive Vitamin E: Why Tocotrienol Trumps Tocopherol

Presented by

Barrie Tan, Ph.D. American River Nutrition, Inc.© January 27, 2009

© 1/09

Vitamin E Perspective Today

• Superstar? • Under-dosing?

• Shocking!

•

Vitamin E – Antioxidant and Nothing More.1 Much higher doses of 3,200IU (2,100mg or 2.1g!) of Vitamin E needed to suppress oxidation in humans.2 95% of Americans do not even have enough of the 12mg RDA, the proclaimed adequacy to prevent a deficiency syndrome.3,4 5

1.

Traber, M.G., and J. Atkinson. (2007) Vitamin E, antioxidant and nothing more. Free Rad Biol Med 43:4 -15.

2.

Robert II, L.J., J.A. Oates, et al. (2007) The relationship between dose of vitamin E and suppression of oxidative stress in humans. Free Rad Biol Med 43(10):138 893.

3.

Maras, J.E., O.I. Bermudez, et al. (2004) Intake of alpha-tocopherol is limited among US adults. J Am Diet Assoc 104(4):567-575.

4.

Traber, M.G. (2006) How much vitamin E?...Just enough! Am J Clin Nutr 84(5):959 -960.

5.

Tocotrienols: Vitamin E beyond Tocopherols. 2008. Eds. R. Watson, V. Preedy. AOCS/CRC Press: Champaign, IL.

5

© 1/09

Vitamin E Vitamin E Family

©1995-2003 by Michael W. Davidson

Tocopherol (T) “Birth Vitamin” (1922)

Delta-T

RDA for Red Blood Cell Function (1960)

Gamma-T Beta-T Alpha-T Most common in vitamin E supplements

6

© 1/09

Vitamin E Vitamin E Family

Tocopherol (T)

Tocotrienol (T3)

Delta-T Gamma-T Beta-T Alpha-T

©1995-2003 by Michael W. Davidson

Delta-T3 Gamma-T3 Beta-T3 Desmethyl isomers Most potent in supporting heart health

Alpha-T3 7

© 1/09

Tocotrienol Trumps Tocopherol

• Both tocotrienol and

Chromanol nucleus

Tocopherol

BUT…. • Only tocotrienol has been shown to: – reduce cholesterol – manage diabetes – inhibit cancer

Tocotrienol

tocopherol are antioxidants

Tocopherol Tocopherol

Tocotrienol Tocotrienol

Shorter farnesyl tail

8

© 1/09

Tail Discussion (1) • Shorter tail = less anchor = move faster = greater reach to all membranes • Trienol Tail – Flip-flops less – Waggles less – Wobbles less

• Conclusion: Shorter tail moves faster & better to protect a much larger cell membrane area (50x better) 9

9

© 1/09

Head Discussion (2) • Less stuffed head (less methyl groups) = smaller head (desmethyl head) = access to action • Desmethyl Head – Protracts (bobs and floats) – Recharges faster – Treat damage faster

• Conclusion: Smaller head arrests/seals damaged membrane efficiently (50x better) 10 * Packer, L., S. U. Weber, et al. (2001). "Molecular aspects of alpha-tocotrienol antioxidant action and cell signalling." J Nutr 131(2): 369S-73S. * Atkinson, J., R. F. Epand, et al. (2008). "Tocopherols and tocotrienols in membranes: a critical review." Free Radic Biol Med 44(5): 739-64.

© 1/09

Desmethyl Tocotrienols

• Desmethyl or less methylated (most potent; Mills-Nixon Effect*) – Delta-T3 and gamma-T3

• Fully or C-5 methylated (less or non-potent) – Alpha-T3 and beta-T3 δT3

γT3

* Behan, J.M., F.M. Dean, and R.A.W. Johnstone. 1976. Photoelectron spectra of cyclic aromatic ethers. Tetrahedron 32:167-171.

11

© 1/09

Interferences by Alpha-Tocopherol • Blocks absorption of tocotrienols1 • Compromises tocotrienol’s ability to reduce cholesterol2,5 • Induces breakdown of tocotrienols3 • Increases cholesterol in high doses4,5 1.

Ikeda, S., et al. 2003. Dietary alpha-tocopherol decreases alpha-tocotrienol but not gamma-tocotrienol concentration in rats. J Nutr 133:428-434.

2.

Qureshi, A. A., et al. 1996. Tocopherol attenuates the impact of gamma-tocotrienol on HMG-CoA reductase activity in chickens. J Nutr 126:389-394.

3.

Sontag, T.J., and R.A. Parker. 2007. Comparative influence of major structural features of tocopherols and tocotrienols on kinetics of their ω -oxidation by cellular and microsomal tocopherol-ω-hydroxylase. J Lip Res 48(5):1090-8.

4.

Stocker, A. 2004. Molecular mechanisms of vitamin E transport. Ann NY Acad Sci 1031:44-59.

5.

Khor, H. T. and T. T. Ng 2000. Effects of administration of alpha-tocopherol and tocotrienols on serum lipids and liver HMG CoA reductase activity. Int J Food Sci Nutr 51 Suppl: S3-11.

12

© 1/09

Alpha-Tocopherol Attenuates Impact of Desmethyl Tocotrienol αT3 < 15% γT3 δT3 > 60% aT1 < 15%+and gT3and + dT3 > 60%

HMGR

Cholesterol

aT1 > 30%+and gT3and + dT3 < 45% αT3 > 30% γT3 δT3 < 45%

HMGR?

Cholesterol?

• Effective preparation for cholesterol reduction: <15% α-tocopherol, >60% desmethyl tocotrienols

• Less effective or ineffective preparation: >30% a-tocopherol, <45% desmethyl tocotrienols

• Shown in numerous animal studies (hamster, guinea pig, rat, chicken) with alpha-T by itself or in combo with tocotrienols * Qureshi, A. A., B. C. Pearce, R. M. Nor, A. Gapor, D. M. Peterson, and C. E. Elson. 1996. Dietary alpha-tocopherol attenuates the impact of gamma-tocotrienol on hepatic 3-hydroxy-3methylglutaryl coenzyme A reductase activity in chickens. J Nutr 126:389-94. * Khor, H. T. and T. T. Ng 2000. Effects of administration of alpha-tocopherol and tocotrienols on serum lipids and liver HMG CoA reductase activity. Int J Food Sci Nutr 51 Suppl: S3-11. 13 * Khor, H.T. and D.Y. Chieng. 1997. Lipidemic effects of tocotrienols, tocopherols and squalene: studies in hamster. Asia Pac J Clin Nutr 6(1):36-40. * Chen, H.W., C.K. Lii, et al. 1995. Dietary fat and vitamin E have differential effects on serum lipid levels. Nutr Res 15:1367-1376.

© 1/09

Diet and Supplement Vitamin E

Dietary Vitamin E

Supplement Vitamin E

* Tan, B. 2005. Appropriate spectrum vitamin E and new perspectives on desmethyl tocopherols and tocotrienols. JANA 8(1):35-42. 14 * Eitenmiller, R. and J. Lee. 2004. Vitamin E: food chemistry, composition and analysis. New York, Marcel Dekker: 425-505.

Annatto Tocotrienols

© 1/09

Rice

Palm

Annatto

Tocopherol

30 – 40% Des T3

Tocotrienol

Tocotrienol

50 - 55% Des T3

> 99% Des T3 Figure 1

100

100

100

80

80

80

60

60

60

40

40

40

20

20

20

0

Gamma

Delta

0

Gamma

Delta

0

Gamma

% Tocopherol

Tocotrienol Tocopherol

Delta

• Only desmethyl Vitamin Es • Exclusively delta-T3 and gamma-T3 • Tocopherol-free • All-natural

Palm and rice have 150-300x more tocopherols than annatto. 15

© 1/09

Bixa orellana: Annatto •

Natural source of tocotrienols from annatto beans

• 30-60x more potent

• The very best vitamin E tocotrienol nature makes! 16

© 1/09

Cholesterol Reduction Timeline (1) 1980 – Discovery of tocotrienol’s hypocholesterolemic activity (U. Wisconsin/Madison)

1992 – Hypocholesterolemic Activity of Tocotrienols Specified (Bristol-Myers Squibb) • • • •

Delta- and gamma-tocotrienols are most active Alpha-tocotrienol is 5-30 times less active Alpha-tocopherol is inactive When combined, only delta-tocotrienol and gamma-tocotrienol work best synergistically

1996 – Discovery of alpha-tocopherol’s interference with tocotrienol’s cholesterol-lowering effects 17

© 1/09

Cholesterol Reduction Timeline (2) 2006 – Dose-Response Impact of Tocotrienols in Animals • Which tocotrienol works better? • Delta-tocotrienol worked best, followed by gamma-tocotrienol • Potency of cholesterol-reduction is delta > gamma > TRF > alpha • Alpha-tocopherol is inactive 2006 – Cholesterol-Reducing Mechanism of Tocotrienol Confirmed (U. Texas) • Unequivocally confirms original Bristol-Myers Squibb study of 1992 • Tocotrienol blocks and degrades the HMGR enzyme protein • Only delta- and gamma-tocotrienol decreases HMGR reductase effectively, while alpha-tocotrienol is 10-fold less active • Only delta-tocotrienol blocks HMGR • Alpha-tocopherol does not work • Endorsed/acknowledged by 1985 Nobel Prize winners for the 18 discovery of the LDL receptor, Goldstein & Brown

Mechanism of Action

© 1/09

Acetyl CoA

HMG CoA Reductase

Statin

(HMGR)

(targets HMGR) Statin inhibits entire IP

Degrades

Downregulates

Isoprenoid Pool (IP) Geraniol Farnesol Geranylgeraniol Cholesterol

Tocotrienol

•Hypercholesterolemia  •Lipidemia 

(targets HMGR) Proteins Protein  by statin may lead to: •Global Myopathy •Anemia

CoQ10 CoQ10  by statin may lead to: •Congestive Heartfailure & Chronic Myopathy •Chronic Fatigue Syndrome & Energy/ATP 

*Pearce, B. C., R. A. Parker, M. E. Deason, A. A. Qureshi, and J. J. Wright. 1992. Hypocholesterolemic activity of synthetic and natural tocotrienols. J Med Chem 35:3595-606. *Song, B. L., and R. A. DeBose-Boyd. 2006. Insig-dependent ubiquitination and degradation of 3-hydroxy-3-methylglutaryl coenzyme a reductase 19 stimulated by delta- and gamma-tocotrienols. J Biol Chem 281:25054-61.

© 1/09

1992 Clinical – University of Wisconsin, Madison Findings with TRF (200mg/day): – Total cholesterol reduction: 15-22% – LDL cholesterol reduction: 10-20% – Of which 15% of patients did not respond to supplementation Additional supplementation with 100mg/day delta- and gamma-tocotrienol

Total cholesterol drop of 35 – 40% following 4 weeks supplementation

20

* Qureshi N., A. A. Qureshi. (1993). Tocotrienols, novel hypocholesterolemic agents with antioxidant properties. Vitamin E in Health and Disease. J. F. L. Packer. New York, Mercel Decker, Inc.: 247-267.

© 1/09

1993 Clinical – BMS-AMR 4-Week Internal Study

Findings with Delta- and Gamma-Tocotrienol (100mg/day):

Gamma-Tocotrienol 10

% Change

– Total cholesterol and LDL reduction: 20-25% – Triglyceride reduction: 15-20% – Delta-tocotrienol ≥ Gamma-tocotrienol

20

Effects of Delta- and Gamma-tocotrienols (100mg/day for 4 wks.) on Lipid Parameters

Delta-Tocotrienol

0

-10

-20

-30 Serum Cholesterol

LDL

HDL

TG

Lipid Parameters

21

© 1/09

2002 Clinical – Dose-Dependent CholesterolReduction by TRF in Hypercholesterolemic Humans

Findings of dosedependent study: – Optimal dose is 75100mg/day – Total cholesterol and LDL reduction: 15-20% – Triglyceride reduction: 7-10%

*Qureshi, A. A., S. A. Sami, W. A. Salser, and F. A. Khan. 2002. Dose-dependent suppression of serum cholesterol by tocotrienol-rich fraction (TRF25) of rice bran in hypercholesterolemic humans. Atherosclerosis 161:199-207. 22

2004 Clinical – Lipid Profile of Hypercholesterolemic Patients on Annatto Tocotrienols

Findings of Tocopherol-Free Delta- and GammaTocotrienol (75mg/day): – Total cholesterol & LDL: 15%↓ – Triglyceride: 20-25%↓ – Cardiovascular risk (TC/HDL): 15-20%↓ – Metabolic syndrome ratio (TG/HDL): 20-30%↓ – Plasma CoQ10: 20%↑ (equi. to 30mg CoQ suppl.)

10

0

%Change

© 1/09

-10

-20

Study A -30

TC

LDL

Study B HDL

TG

TC

LDL

HDL

TG

Lipid Parameters

* Tan, B. and A. M. Mueller (2008). Tocotrienols in Cardiometabolic Diseases. Tocotrienols: Vitamin E beyond Tocopherol. R. Watson, V. Preedy, CRC/AOCS Press: 257-273.

23

© 1/09

Atherosclerosis • Athere (Gruel, thickening) - stroke, heart attack • Skleros (Narrowing) - high blood pressure, small vessel problems

From Time: Your Body, A User’s Guide, 2008

• Atherosclerosis - hardening & blocking of arteries, resulting in arterial dysfunction 24

© 1/09

Tocotrienol Reduces Plaque and Plaque Rupture

Atherogenic Diet

Cholesterol Crystals

Atherogenic Diet and Tocotrienol

Protruding cholesterol crystals at plaque rupture site of human coronary artery

*Abela GS, and K Aziz. 2005. Cholesterol crystals cause mechanical damage to biological membranes: A proposed mechanism of plaque rupture and erosion leading to arterial thrombosis. Clin Cardiol 28:413420. 25 *Abela, G.S., and K. Aziz. 2006. Cholesterol crystals rupture biological membranes and human plaques during acute cardiovascular events – A novel insight into plaque rupture by SEM. Scanning 28(1):1-10. * Black et al. 2000. Palm tocotrienols protect apoE +/- mice from diet-induced atheroma formation. J Nutr 130:2420-2426.

© 1/09

Tocotrienol Reduces Carotid Atherosclerosis • Tocotrienols regressed carotid artery stenosis in humans in a 4-year study • With T3: 88% of subjects either improved or stabilized • Without T3: 60% of subjects deteriorated

26

* Kooyenga, D. K., T. R. Watson, et al. (2001). Antioxidants modulate the course of carotid atherosclerosis: A four-year report. Micronutrients and Health. K. Nesaretnam and L. Packer. Illinois, AOCS Press: 366-375.

© 1/09

Tocotrienols Inhibit Atherosclerotic Lesions

Diets (Months)

0% DMT3† (Alpha-T)

71% DMT3

100% DMT3

Low Fat (3.5 months)

11%

42%

47%

High Fat (4.5 months)

19%

28%

33%

High Fat (6 months)

23%

36%

57%

†DMT3

= Delta- & gamma-tocotrienol

* Qureshi, AA, et al. 2001. Novel tocotrienols of rice bran inhibit atherosclerotic lesions in C57BL/6 apoE-deficient mice. J Nutr 131:2606-2618. * Black, T. M., P. Wang, et al. (2000). "Palm tocotrienols protect ApoE +/- mice from diet-induced atheroma formation." J Nutr 130: 2420-2426.

27

© 1/09

Dosage and Duration in Clinical Studies

Study (Source)

Dosage (mg/d)

Duration (months)

LDL ↓ (%) Triglyceride ↓ (%)

Hypercholesterolemia (Palm)

500

1

15

13

Carotid Atherosclerosis (Palm & Rice)

200-350

48

14

-

Hypercholesterolemia (Palm*)

200

1.5

0

-

Hypercholesterolemia (Rice†)

100

1

20

8

Dyslipidemia (Pure Delta-T3, Gamma-T3)

100

1

20-25

15-20

Dyslipidemia (Annatto)

75

2

15

20-25

* 33% alpha-tocopherol † < 10% alpha-tocopherol and > 80% tocotrienols 1. 2. 3. 4. 5. 6.

Ajuluchukwu, J. N., N. U. Okubadejo, et al. (2007). "Comparative study of the effect of tocotrienols and -tocopherol on fasting serum lipid profiles in patients with mild hypercholesterolaemia: a preliminary report." Niger Postgrad Med J 14(1): 30-3. Kooyenga, D. K., T. R. Watson, et al. (2001). Antioxidants modulate the course of carotid atherosclerosis: A four-year report. Micronutrients and Health. K. Nesaretnam and L. Packer. Illinois, AOCS Press: 366-375. Qureshi, A.A., D.A. Khan, et. Al. Interaction of alpha-tocopherol with palmvitee and lovastatin in cholesterol modulation in hypercholesterolemic human subjects. Manuscript in preparation. Qureshi, A. A., S. A. Sami, W. A. Salser, and F. A. Khan. (2002). Dose-dependent suppression of serum cholesterol by tocotrienol-rich fraction (TRF25) of rice bran in hypercholesterolemic humans. Atherosclerosis 161:199-207. BMS-AMR study (slide #19) 28 Tan, B. and A. M. Mueller (2008). Tocotrienols in Cardiometabolic Diseases. Tocotrienols: Vitamin E beyond Tocopherol. R. Watson, V. Preedy, AOCS/CRC Press: 257-273.

© 1/09

• •

Effect of Tocopherol and Tocotrienol on Platelet Aggregation (PA) in Stenosed Coronary Canine Artery

Tocopherol-free tocotrienol (85% Delta & Gamma) significantly reduced PA in dogs with induced coronary atherosclerosis Potency of PA Inhibition: DesT3 (Delta & Gamma) > Alpha-T3 > AlphaT

29

* Qureshi, A.A., C.W. Karpen, et al. (2009). “Tocotrienol-induced inhibition of platelet thrombus formation and platelet aggregation in a stenosed coronary artery canine model.” Manuscript in preparation.

© 1/09

Diabetes by the Numbers

60 million Pre-diabetics

27 million Diabetics in 2005, about one quarter did not know

18.2 million Diabetics in 2003 0

10

20

30

40

50

60

70

30

© 1/09

Complications of Diabetes • 2-4 times higher risk of heart disease and stroke • 75% of adults with diabetes have HBP • 60-70% of people with diabetes have nerve-artery damage • Leading cause of blindness and kidney failure

31

© 1/09

AHA and NIH Metabolic Syndrome Recommendations • Metabolic syndrome is identified by presence of three or more of these risk factors: – Elevated waist circumference – Elevated triglycerides – Elevated blood pressure – Elevated fasting glucose – Reduced HDL cholesterol

32

© 1/09

Tocotrienol’s Effect in Type 2 Diabetes • T3 decreases triglyceride levels • T3 reduces bad LDL and increases good HDL • The sum: Delta-T3 and Gamma-T3 may help manage diabetes and prediabetes • T3 reduces symptoms of diabetes in animals: high blood pressure, arterial integrity 33 33

2008 Leading Cancer Killers*

© 1/09

Women

Men Lung (90,810)

Lung (71,030)

Breast (40,480)

Prostate (28,660)

Colon and rectum (24,260)

Colon & Rectum (25, 700)

Pancreas (16,790)

Pancreas (17,500)

Ovary (15,520)

Liver (12,570)

Non-Hodgkin lymphoma (9,370)

Leukemia (12,460)

Leukemia (9,250)

Esophagus (11,250)

Uterus (7,470)

Bladder (9,950)

Liver (5,840) 80,000

70,000

60,000

50,000

40,000

30,000

20,000

10,000

*Source: American Cancer Society, Cancer Facts & Figures 2008.

Non-Hodgkin Lymphoma (9,790) 0

0

10,000

20,000

30,000

40,000

50,000

60,000

70,000

80,000

90,000

34

100,000

2008 Leading Cancer Killers*

© 1/09

Women

Men Lung (90,810)

Lung (71,030)

Breast (40,480)

Prostate (28,660)

Colon and rectum (24,260)

Colon & Rectum (25, 700)

Pancreas (16,790)

Pancreas (17,500)

Ovary (15,520)

Liver (12,570)

Non-Hodgkin lymphoma (9,370)

Leukemia (12,460)

Leukemia (9,250)

Esophagus (11,250)

Uterus (7,470)

Bladder (9,950)

Liver (5,840) 80,000

70,000

60,000

50,000

40,000

30,000

20,000

10,000

*Source: American Cancer Society, Cancer Facts & Figures 2008.

Non-Hodgkin Lymphoma (9,790) 0

0

10,000

20,000

30,000

40,000

50,000

60,000

70,000

80,000

90,000

35

100,000

© 1/09

Tocotrienols More Potent than Tocopherols In Vivo and In Vitro 30 – 60x more potent

• Desmethyl tocotrienols dramatically increased bioavailability • Inhibiting adhesion molecules, suppressing cancer growth, accumulating in tumors:

δT3 > γT3 > αT3 > δT > γT > αT *Naito, Y., M. Shimozawa, M. Kuroda, N. Nakabe, H. Manabe, K. Katada, S. Kokura, H. Ichikawa, N. Yoshida, N. Noguchi, and T. Yoshikawa. 2005. Tocotrienols reduce 25-hydroxycholesterol-induced monocyteendothelial cell interaction by inhibiting the surface expression of adhesion molecules. Atherosclerosis 180:19-25. 36 *McIntyre, B. S., K. P. Briski, M. A. Tirmenstein, M. W. Fariss, A. Gapor, and P. W. Sylvester. 2000. Antiproliferative and apoptotic effects of tocopherols and tocotrienols on normal mouse mammary epithelial cells. Lipids 35:171-80. * Hiura, Y., H. Tachibana, et al. 2008. Specific accumulation of gamma- and delta-tocotrienols in tumor and their antitumor effect in vivo. J Nutr Biochem. In press 2009.

© 1/09

Delta-Tocotrienol Treatment and Prevention of Human Pancreatic Cancer*

• Apoptosis against pancreatic cancer cells – Delta-T3 >> gemcitabine, gamma-T3 – Delta-T3 is most potent – Delta-T3 is non-toxic to nontransformed cells

• Preferred composition

Control

Delta-T3

– Delta-T3 and/or Gamma-T3 – Free of Alpha-T3, Beta-T3 – Free of tocopherol 37

* Malafa, M. P. and S. Sebti (2008). Delta-Tocotrienol Treatment and Prevention of Pancreatic Cancer. US, Lee Moffitt Cancer Center & Research Institute, University of South Florida (Tampa). * Husain, K., R. Francois, et. Al. 2008. Delta-tocotrienol is the most bioactive natural tocotrienol in the prevention of pancreatic cancer transformation. Am Assoc Canc Res (April 12-16, 2008) San Diego, Abstr 3826.

© 1/09

Anti-Angiogenic Properties of Tocotrienols (1)

• Angiogenesis is important in tumor growth, diabetic retinopathy, rheumatoid arthritis, and psoriasis • Delta-T3 is most active; without toxicity • Alpha-T does not work • May work better with omega3, resveratrol, EGCG, capsaicin • Possible mechanisms: – VEGF, FGF, EGF – Telomerase – Hypoxia * Nakagawa, K., T. Eitsuka, et al. (2004). "DNA chip analysis of comprehensive food function: inhibition of angiogenesis and telomerase activity with unsaturated vitamin E, tocotrienol." Biofactors 21(1-4): 5-10. * Mizushina, Y., K. Nakagawa, et al. (2006). "Inhibitory effect of tocotrienol on eukaryotic DNA polymerase lambda and angiogenesis." Biochem Biophys Res Commun 339(3): 949-55. 38 * Shibata, A., K. Nakagawa, et al. (2008). "Tocotrienol inhibits secretion of angiogenic factors from human colorectal adenocarcinoma cells by suppressing hypoxia-inducible factor-1alpha." J Nutr 138(11): 2136-42.

© 1/09

Anti-Angiogenic Properties of Tocotrienols (2)

• In tumor-implanted mice: (A) Without Delta-T3 – Neovascularization (B) With Delta-T3 – Inhibition of neovessel formation

• Anti-angiogenic effect of Delta-T3 attributable to

Neovascularization

– Inhibition of growth factors (e.g. vascular endothelial, fibroblast, epidermal) – Generation of ROS and apoptosis in tumor

* Shibata, A., K. Nakagawa, et al. (2008). "Tumor anti-angiogenic effect and mechanism of action of delta-tocotrienol." Biochem Pharmacol 76(3): 330-9. * Miyazawa, T., A. Shibata, et al. (2009). "Antiangiogenic and anticancer potential of unsaturated vitamin E (tocotrienol)." J Nutr Biochem 20(2): 79-86.

39

Delta-Tocotrienol Excels in Inducing Growth Inhibition of Mammary Cancer Cells

© 1/09

>120uM 100 90

Potency to induce growth inhibition of mammary tumor cells by 50%:

70

δT3 > γT3 > αT3 > TRF > δT >> αT

60

Preneoplastic

Gamma-T3 Delta-T3

TRF

0

TRF

10

TRF

20

More invasive mammary cancer, more responsive to tocotrienols:

Gamma-T3 Delta-T3

30

Gamma-T3 Delta-T3

40

Alpha-T

Alpha-T

50

Alpha-T

Treatment Dose to Induce IC 50 (uM)

80

δT3 > γT3 > αT3 > TRF > δT >> αT

Neoplastic Malignant * McIntyre, B. S., K. P. Briski, et al. 2000. Antiproliferative and apoptotic effects of tocopherols and tocotrienols on preneoplastic and neoplastic mouse mammary e pithelial cells. Proc Soc Exp 40 Biol Med 224(4): 292-301. * Sylvester, P. W. and S. J. Shah. 2005. "Mechanisms mediating the antiproliferative and apoptotic effects of vitamin E in mammary cancer cells." Front Biosci 10: 699-709.

© 1/09

Interferences by Alpha-Tocopherol in Mammary Cancer Cells • Interferes with estrogen (+ve & –ve) cancer • Antagonizes tamoxifen’s ability to block estrogen +ve cancer by 1,000x • Blocks completely the powerful effects of tamoxifen on estrogen –ve cancer • Argues for tocopherol-free desmethyl tocotrienol (delta-T3 & gamma-T3) usage

1.

Schwenke, D. C. (2002). "Does lack of tocopherols and tocotrienols put women at increased risk of breast cancer?" J Nutr Biochem 13(1): 2-20.

2.

Gundimeda, U., Z. H. Chen, et al. (1996). "Tamoxifen modulates protein kinase C via oxidative stress in estrogen receptor-negative breast cancer cells." J Biol Chem 271(23): 13504-14.

3.

Guthrie, N., A. Gapor, et al. (1997). "Inhibition of proliferation of estrogen receptor-negative MDA-MB-435 and -positive MCF-7 human breast cancer cells by palm oil tocotrienols and tamoxifen, alone and in combination." J Nutr 127: 544S-548S.

41

Tocotrienol and Melanoma

• Desmethyl tocotrienols suppress tumor growth in vitro and in vivo • Via suppression of HMG CoA reductase activity • Tumor suppression: – delta-T3: 50% – delta-T3 + Lov: 60%

• Combo is nontoxic to animals at tumor-suppressive doses with T3:Lov of 5:1 • T3+Lov combo has already been successfully shown in hypercholesterolemic patients

100 90 80 T umor Weight (%)

© 1/09

70

Desmethyl isomers

60 50 40 30 20 10 0 Control

Alpha-T

Gamma-T3

Delta-T3

924

630

Delta-T3 + Lov Blend

u mol/kg

*He, L., H. Mo, S. Hadisusilo, A. A. Qureshi, and C. E. Elson. 1997. Isoprenoids suppress the growth of murine B16 melanomas in vitro and in vivo. J Nutr 127:668-74. *McAnally J.A., J. Gupta, S. Sodhani, L. Bravo, H. Mo. 2007. Tocotrienols potentiate lovastating-mediated growth suppression in vitro and in vivo. Exp Biol Med 232(4):523-31.

42

*Qureshi, A. A., S. A. Sami, et al. 2001. Synergistic effect of tocotrienol-rich fraction (TRF(25)) of rice bran and lovastatin on lipid parameters in hypercholesterolemic humans. J Nutr Biochem 12(6): 318-329.

© 1/09

Multifaceted Mechanisms of Tocotrienol

• Much higher cellular and tumor bioavailability than tocopherol (50100x), especially delta-T3 and gamma-T3 • HMG CoA reductase suppression • Pro-apoptotic effects • Anti-angiogenesis (growth factors, hypoxia) • Genetic regulations

Foundation for Cancer Research and Education

* Hiura, Y., H. Tachibana, et al. (2008). "Specific accumulation of gamma- and delta-tocotrienols in tumor and their antitumor effect in vivo." J Nutr Biochem. 43 * Mo, H. and C. E. Elson (2004). "Studies of the isoprenoid-mediated inhibition of mevalonate synthesis applied to cancer chemotherapy and chemoprevention." Exp Biol Med (229): 567-585. * Miyazawa, T., A. Shibata, et al. (2009). "Antiangiogenic and anticancer potential of unsaturated vitamin E (tocotrienol)." J Nutr Biochem 20(2): 79-86.

© 1/09

Antioxidant Values of Various Oils

DeltaGold® may be used as a food protectant (~500ppm) or as a super-antioxidant in combination with EGCG and resveratrol. * Kim, H. J. 2007. Oxidation mechanism of riboflavin destruction and antioxidant mechanism of tocotrienols. Food Science and Nutrition. Columbus, The Ohio State University. Ph.D.

*Qureshi, A. A., H. Mo, L. Packer, and D. M. Peterson. 2000. Isolation and identification of novel tocotrienols from rice bran with hypocholesterolemic, antioxidant, and antitumor properties. J Agric Food Chem 48:313040. 44 *Chin, S.F., N.A. Hamid, et al. 2008. Reduction of DNA damage in older healthy adults by Tri E((R)) Tocotrienol supplementation. Nutrition 24(1):1-10. *Yu, F. L., A. Gapor, and W. Bender. 2005. Evidence for the preventive effect of the polyunsaturated phytol side chain in tocotrienols on 17beta-estradiol epoxidation. Cancer Detect Prev 29:383-8.

© 1/09

Antioxidation Action of Tocotrienol • Oxidized LDL • Advanced glycation end (AGE) products • Glycated hemoglobin (HbAlc) • Damaged DNAs • Adhesion molecules • Platelet aggregation • Thromboxanes • Leukotrienes • CRP, TNF, NFkB

* Kim, H. J. 2007. Oxidation mechanism of riboflavin destruction and antioxidant mechanism of tocotrienols. Food Science and Nutrition. Columbus, The Ohio State University. Ph.D. 45 *Kim, H.J., and D.B. Min. 2007. Effects, quenching mechanisms, and kinetics of alpha-, beta-, gamma-, and delta-tocotrienol on chlorophyll photosynthesized oxidation of lard. Presented at IFT Conference, July 2007.

© 1/09

Product Concepts

1. T3 + (Sterol/RRY) * Healthy lipids (cholesterol) * Inside-Outside program (T3Sterol) * One-Two punch (T3-RRY) 2. T3 + Omega 3 * Healthy lipids (triglycerides) * Healthy arteries * Healthy RBCs

3. T3 + (EGCG/Resveratrol) * Healthy arteries * Healthy cells * Super lipid antioxidants 4. T3 + Lignans (Sesame/ Flaxseed) * Potentiate T3 * Increase blood levels

* Duncan, R.E., A. El-Sohemy, and M.C. Archer. 2005. Regulation of HMG-CoA reductase in MCF-7 cells by genistein, EPA and DHA, alone and in combination with mevastatin. Cancer Letters 224:221-228. * McAnally, J.A., J. Gupta, S. Shodhani, L. Bravo, and H.B. Mo. 2007. Tocotrienols potentiate lovastatin-mediated growth suppression in vitro and in vivo. Exp Biol Med 232:523-531. * Tan, B., and J. Llobrera. February 17, 2005. Annatto extract compositions including tocotrienols and tocopherols and methods of use. patent 2005003710 2. 46

© 1/09

Eye Formula • • • • • •

1. 2. 3. 4. 5. 6. 7.

Dry eye1 Diabetic Retinopathy2 Retinopathy of Prematurity3 Glaucoma4 Cornea (cataract)5 T3 + Ω3 combo6,7

Rubin, B. Y., S. L. Anderson, et al. (2008). "Can the therapeutic efficacy of tocotrienols in neurodegenerative familial dysa utonomia patients be measured clinically?" Antioxid Redox Signal 10(4): 837-41. Miyazawa, T., A. Shibata, et al. (2008). "Anti-angiogenic function of tocotrienol." Asia Pac J Clin Nutr 17 Suppl 1: 253-6. Hittner, H., F.L. Kretzer. (1986). “Efficacy of Vitamin E in Retinopathy of Prematurity.” Retinopathy of Prematurity: Current Concenpts and Controversies. A.R. McPherson, H.M. Hittner, F.L. Kretzer. B.C. Decker Inc: 89-103. Meyenberg, A., D. Goldblum, et al. (2005). "Tocotrienol inhibits proliferation of human Tenon's fibroblasts in vitro: a compa rative study with vitamin E forms and mitomycin C." Graefes Arch Clin Exp Ophthalmol 243(12): 1263-71. Tanito, M., N. Itoh, et al. (2004). "Distribution of tocopherols and tocotrienols to rat ocular tissues after topical ophthal mic administration." Lipids 39(5): 469-74. 47 Miljanovic, B., K. A. Trivedi, et al. (2005). "Relation between dietary n-3 and n-6 fatty acids and clinically diagnosed dry eye syndrome in women." Am J Clin Nutr 82(4): 887-93. Rashid, S., Y. Jin, et al. (2008). "Topical omega-3 and omega-6 fatty acids for treatment of dry eye." Arch Ophthalmol 126(2): 219-25.

© 1/09

Tocotrienol-CoQ10 Formula • CoQ10 and Systolic BP ↓ (6-18mm Hg)1-3 • CoQ10 and Diastolic BP ↓ (3-10 mm Hg)

• T3 and NOS ↑, Lipid Perox. ↓, SH ↓4-5 • T3 and Systolic BP ↓ (3-6 mm Hg)6 • T3 and arterial integrity ↑7 • Combo Concept per day: 100-200mg CoQ10 100mg Tocotrienol 1.

Burke, B.E., et al. 2001. Randomized, double-blind, placebo-controlled trial of coenzyme Q10 in isolated systolic hypertension. South Med J 94(11):1112-7.

2.

Rosenfeldt, F.L., et al. 2007. Coenzyme Q10 in the treatment of hypertension: a meta-analysis of the clinical trials. J Hum Hypertens 21:297-306.

3.

Hodgson J.M., et al. 2002. Coenzyme Q10 improves blood pressure and glycaemic control: a controlled trial in subjects with type 2 diabetes. Eur J Clin Nutr 56(11):1137-42.

4.

Newaz, M. A., and N. N. Nawal. 1999. Effect of gamma-tocotrienol on blood pressure, lipid peroxidation and total antioxidant status in spontaneously hypertensive rats (SHR). Clin Exp Hypertens 21:1297-313.

5.

Newaz, M. A., Z. Yousefipour, N. Nawal, and N. Adeeb. 2003. Nitric oxide synthase activity in blood vessels of spontaneously hypertensive rats: antioxidant protection by gamma-tocotrienol. J Physiol Pharmacol 54:319-27.

6.

Rasool, A.H.G., K.H. Yuen, K. Yusoff, A.R. Wong, and A.R.A. Rahman. 2006. Dosepdependent elevation of plasma tocotrienol levels and its effect on arterial compliance, plasma total antioxidant 48 status, and lipid profile in healthy humans supplemented with tocotrienol -rich vitamin E. J Nutr Sci Vitaminol 52:473-478.

7.

Rasool, A. H., A. R. Rahman, et al. 2008. Arterial compliance and vitamin E blood levels with a self emulsifying preparation of tocotrienol rich vitamin E. Arch Pharm Res 31(9): 1212-7.

© 1/09

Conclusion

• DeltaGold® Annatto Tocotrienol is/has – – – – –

Multi-patent protection Tocopherol-free Exclusively Delta-T3 and Gamma-T3 Best-in-Class Tocotrienol Made in the USA

• DeltaGold® Availability – 50% Oil – 35% Powder

49

© 1/09

First-Ever Tocotrienol Book!

This and more literature on vitamin E tocotrienol available on our website, www.AmericanRiverNutrition.com

50

© 1/09

Contact Us! DeltaGold® Tocotrienol Your only source of tocopherol-free tocotrienol [email protected] www.AmericanRiverNutrition.com (413) 253-3449

American River Nutrition, Inc. ∙ 31 Campus Plaza Rd. ∙ Hadley, MA 01035

51