Decoding the Elusive Vitamin E: Why Tocotrienol Trumps Tocopherol

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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

Presented by

Barrie Tan, Ph.D.American River Nutrition, Inc.©

January 27, 2009

Decoding the Elusive Vitamin E: Why Tocotrienol Trumps Tocopherol

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5

• Superstar? Vitamin E – Antioxidant and Nothing More.1

• Under-dosing? Much higher doses of 3,200IU (2,100mg or 2.1g!) of Vitamin E needed to suppress oxidation in humans.2

• Shocking! 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 8-93.

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.

Vitamin E Perspective Today

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6

Vitamin E

Vitamin E Family

Tocopherol (T)

Alpha-T

Beta-T

Gamma-T

Delta-T

Most common in vitamin E supplements

“Birth Vitamin” (1922)

RDA for Red Blood Cell Function (1960)

©1995-2003 by Michael W. Davidson

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7

Vitamin E Family

Tocopherol (T) Tocotrienol (T3)

Alpha-T

Beta-T

Gamma-T

Delta-T Delta-T3

Gamma-T3

Beta-T3

Alpha-T3Desmethyl isomers -Most potent in supporting heart health

Vitamin E

©1995-2003 by Michael W. Davidson

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8

Tocopherol Tocotrienol

Tocotrienol Trumps Tocopherol

• Both tocotrienol and tocopherol are antioxidants

Tocopherol Tocotrienol

Chromanol nucleus

• Only tocotrienol has

been shown to:

– reduce cholesterol

– manage diabetes

– inhibit cancer

BUT….

Shorter farnesyl tail

Toco

ph

ero

l

Toco

trie

no

l

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9

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

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10

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)

* 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.

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11

Desmethyl Tocotrienols

δT3 γT3

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

• 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

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12

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 JFood Sci Nutr 51 Suppl: S3-11.

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13

Alpha-Tocopherol Attenuates Impact of Desmethyl Tocotrienol

• 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-3-

methylglutaryl 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. * 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.

αT3 < 15% + γT3 and δT3 > 60%

αT3 > 30% + γT3 and δT3 < 45%

HMGR Cholesterol

HMGR? Cholesterol?

aT1 < 15% and gT3 + dT3 > 60%

aT1 > 30% and gT3 + dT3 < 45%

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14

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.* Eitenmiller, R. and J. Lee. 2004. Vitamin E: food chemistry, composition and analysis. New York, Marcel Dekker : 425-505.

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15

Annatto Tocotrienols

• Only desmethyl Vitamin Es

• Exclusively delta-T3 and gamma-T3

• Tocopherol-free

• All-natural

% T

oco

ph

ero

lPalm and rice have 150-300x

more tocopherols than annatto.

0

20

40

60

80

100

Gamma Delta0

20

40

60

80

100

Gamma Delta0

20

40

60

80

100

Gamma Delta

Tocotrienol TocopherolTocotrienol

TocopherolTocotrienol

Rice Palm Annatto

30 – 40% Des T3 50 - 55% Des T3 > 99% Des T3

Figure 1

© 1/09 Bixa orellana: Annatto

• Natural source of tocotrienols from annatto beans

• The very best vitamin E tocotrienol nature makes!

• 30-60x more potent

16

© 1/09

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

Cholesterol Reduction Timeline (1)

17

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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

discovery of the LDL receptor, Goldstein & Brown

Cholesterol Reduction Timeline (2)

18

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19

Mechanism of Action

*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 stimulated by delta- and gamma-tocotrienols. J Biol Chem 281:25054-61.

Acetyl CoA

HMG CoA Reductase

(HMGR)

Cholesterol

Statin

(targets HMGR)

Geraniol

Farnesol

Geranylgeraniol

Isoprenoid Pool (IP)

Tocotrienol

(targets HMGR)

Proteins CoQ10

Statin inhibits

entire IP

•Hypercholesterolemia

•Lipidemia

Protein by statin may lead to:

•Global Myopathy

•Anemia

CoQ10 by statin may lead to:

•Congestive Heartfailure & Chronic Myopathy

•Chronic Fatigue Syndrome & Energy/ATP

De

gra

de

s

Do

wn

reg

ula

tes

© 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

* 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.

20

© 1/09

1993 Clinical – BMS-AMR 4-Week Internal Study

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

– Total cholesterol and LDL reduction: 20-25%

– Triglyceride reduction: 15-20%

– Delta-tocotrienol ≥ Gamma-tocotrienol

20

10

0

-10

-20

-30

Serum

Cholesterol

LDL TGHDL

Delta-Tocotrienol

Gamma-Tocotrienol

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

% C

hang

e

Lipid Parameters

21

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2002 Clinical – Dose-Dependent Cholesterol-Reduction by TRF in Hypercholesterolemic Humans

Findings of dose-dependent study:

– Optimal dose is 75-100mg/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

© 1/09

2004 Clinical – Lipid Profile of HypercholesterolemicPatients on Annatto Tocotrienols

Findings of Tocopherol-Free Delta- and Gamma-Tocotrienol (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.)

Study A Study B

TC LDL HDL TG TC LDL HDL TG-30

-20

-10

0

10

%C

ha

ng

e

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

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24

Atherosclerosis

• Athere(Gruel, thickening)- stroke, heart attack

• Skleros(Narrowing)- high blood pressure, small vessel problems

• Atherosclerosis- hardening & blocking of arteries, resulting in arterial dysfunctionFrom Time: Your Body, A User’s Guide, 2008

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25

Cholesterol Crystals

Tocotrienol Reduces Plaque and Plaque Rupture

*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 28:413-420.*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.

Protruding cholesterol crystals at plaque rupture site of human coronary artery

Atherogenic Diet Atherogenic Diet and Tocotrienol

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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

* 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.

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27

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.

© 1/09

Study (Source) Dosage (mg/d) Duration (months)

LDL ↓ (%) Triglyceride ↓ (%)

Hypercholesterolemia(Palm)

500 1 15 13

Carotid Atherosclerosis(Palm & Rice)

200-350 4 8 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

Dosage and Duration in Clinical Studies

1. 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.

2. 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: 366-375.3. 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.4. 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

161:199-207.5. BMS-AMR study (slide #19)6. 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.

* 33% alpha-tocopherol† < 10% alpha-tocopherol and > 80% tocotrienols

28

© 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* 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.

29

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30

Diabetes by the Numbers

0 10 20 30 40 50 60 70

Pre-diabetics

Diabetics in 2005, about one quarter did not know

Diabetics in 2003

60 million

18.2 million

27 million

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31

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

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32

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

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33

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

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34

2008 Leading Cancer Killers*

0 10,000 20,000 30,000 40,000 50,000 60,000 70,000 80,000 90,000 100,000

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

MenWomen

Lung (90,810)

Liver (12,570)

Bladder (9,950)

Esophagus (11,250)

Leukemia (12,460)

Non-Hodgkin Lymphoma (9,790)

Pancreas (17,500)

Colon and rectum (24,260)

Prostate (28,660)

010,00020,00030,00040,00050,00060,00070,00080,000

Lung (71,030)

Breast (40,480)

Colon & Rectum (25, 700)

Uterus (7,470)

Ovary (15,520)

Liver (5,840)

Pancreas (16,790)

Leukemia (9,250)

Non-Hodgkin lymphoma (9,370)

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35

2008 Leading Cancer Killers*

0 10,000 20,000 30,000 40,000 50,000 60,000 70,000 80,000 90,000 100,000

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

MenWomen

Lung (90,810)

Liver (12,570)

Bladder (9,950)

Esophagus (11,250)

Leukemia (12,460)

Non-Hodgkin Lymphoma (9,790)

Pancreas (17,500)

Colon and rectum (24,260)

Prostate (28,660)

010,00020,00030,00040,00050,00060,00070,00080,000

Lung (71,030)

Breast (40,480)

Colon & Rectum (25, 700)

Uterus (7,470)

Ovary (15,520)

Liver (5,840)

Pancreas (16,790)

Leukemia (9,250)

Non-Hodgkin lymphoma (9,370)

© 1/09

• 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 monocyte-endothelial cell interaction by inhibiting the surface expression of adhesion molecules. Atherosclerosis 180:19-25.*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.

36

Tocotrienols More Potent than Tocopherols In Vivo and In Vitro

30 – 60x more potent

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37

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 non-transformed cells

• Preferred composition– Delta-T3 and/or Gamma-T3

– Free of Alpha-T3, Beta-T3

– Free of tocopherol

Control

Delta-T3

* 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.

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38

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 omega-3, 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.* 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

– 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.

Neovascularization

39

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40

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

Preneoplastic

Neoplastic

Malignant

0

10

20

30

40

50

60

70

80

90

100

Alp

ha-

T

Alp

ha-T

Alp

ha-T

TR

F

TRF

TRF

Del

ta-T

3

Del

ta-T

3

Del

ta-T

3

Ga

mm

a-T

3

Gam

ma

-T3

Gam

ma

-T3

Tre

atm

en

t D

ose

to

In

du

ce I

C5

0(u

M)

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

δT3 > γT3 > αT3 > TRF > δT >> αT

* McIntyre, B. S., K. P. Briski, et al. 2000. Antiproliferative and apoptotic effects of tocopherols and tocotrienols on preneoplastic and neoplastic mouse mammary epithelial cells. Proc Soc Exp

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.

>120uM

More invasive mammary cancer, more responsive to tocotrienols:

δT3 > γT3 > αT3 > TRF > δT >> αT

© 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

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42

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

0

10

20

30

40

50

60

70

80

90

100

Control Alpha-T Gamma-T3 Delta-T3 Delta-T3 + Lov Blend

Tu

mor

Weig

ht (%

)

*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.

*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.

924 630

umol/kg

Desmethyl isomers

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Multifaceted Mechanisms of Tocotrienol

• Much higher cellular and tumor bioavailability than tocopherol (50-100x), especially delta-T3 and gamma-T3

• HMG CoA reductase suppression

• Pro-apoptotic effects

• Anti-angiogenesis (growth factors, hypoxia)

• Genetic regulations

* 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.

* 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.

Foundation for Cancer Research and Education

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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 48:3130-40. *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.

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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.

*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.

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Product Concepts

1. T3 + (Sterol/RRY)

* Healthy lipids (cholesterol)

* Inside-Outside program (T3-Sterol)

* 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.

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Eye Formula

• Dry eye1

• Diabetic Retinopathy2

• Retinopathy of Prematurity3

• Glaucoma4

• Cornea (cataract)5

• T3 + Ω3 combo6,7

1. Rubin, B. Y., S. L. Anderson, et al. (2008). "Can the therapeutic efficacy of tocotrienols in neurodegenerative familial dysautonomia patients be measured clinically?" Antioxid Redox Signal

10(4): 837-41.2. Miyazawa, T., A. Shibata, et al. (2008). "Anti-angiogenic function of tocotrienol." Asia Pac J Clin Nutr 17 Suppl 1: 253-6.3. 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: 89-103.4. Meyenberg, A., D. Goldblum, et al. (2005). "Tocotrienol inhibits proliferation of human Tenon's fibroblasts in vitro: a comparative study with vitamin E forms and mitomycin C." Graefes

Arch Clin Exp Ophthalmol 243(12): 1263-71.5. Tanito, M., N. Itoh, et al. (2004). "Distribution of tocopherols and tocotrienols to rat ocular tissues after topical ophthalmic administration." Lipids 39(5): 469-74.6. 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): 887-93.7. 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.

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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 CoQ10100mg 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 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.

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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

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First-Ever Tocotrienol Book!

This and more literature on vitamin E tocotrienol available on

our website,

www.AmericanRiverNutrition.com

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Contact Us!

DeltaGold® TocotrienolYour only source of tocopherol-free tocotrienol

arn@american-river.com

www.AmericanRiverNutrition.com

(413) 253-3449

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