Slide 1: Up-regulation of the antioxidant enzymes SOD and catalase in vivo
Up-regulation of the antioxidant enzymes SOD and catalase in vivo: A new approach to antioxidant therapy Joe M. McCord, Ph.D. Lifeline Therapeutics ric
Slide 2: What is Oxidative Stress?
What is Oxidative Stress? Oxidative stress occurs when oxidative balance is upset by increased production of oxidants (O,"", H,O;), or by decreased availability of antioxidants. We normally make about 0.3 mole of O,"“/day, and nearly all is scavenged by our own antioxidant enzymes, the SODs. “Consum
Slide 3: Excess oxidants can attack lipids, proteins and nucleic acids
Excess oxidants can attack lipids, proteins and nucleic acids Lipids are the most readily oxidizable class of biomolecules, so oxidative stress is most often monitored by measuring the products of lipid peroxidation (TBARS, isoprostanes). DNA damage may be measured as 8-OHdG; protein damage may be m
Slide 4: Plasma TBARS increase with age in normal healthy humans
Plasma TBARS increase with age in normal healthy humans © e@ R=0238 @ Females TBARS (micromolar) @ Males
Slide 5: TBARS generated by elevated oxidative stress
TBARS generated by elevated oxidative stress ea _f i a ae \ of Acrolein SWyorel wd 4-Hydi /-2-alkenal: pat “ on a _ arenes Lipid derived OS, ve he “Tr = 8 Malondialdehyde 2Hydroxypropanal— othyigiyoxal HOW 6H HOW way A Carbohydrate derived bi 8 ou 8 Glucosone 3-Deoxyglucosone sor
Slide 6: Modification of LDL to form oxidized LDL
Modification of LDL to form oxidized LDL malondialdehyde °s-\_, + Apolipoprotein B or LDL a oxidized LDL (oxLDL) When >15% of lysine residues of LDL are modified by MDA adducts, the protein is no longer recognized by the LDL receptors in hepatic and peripheral cells, but it is recognized by the scav
Slide 7: The Oxidative Modification Hypothesis
The Oxidative Modification Hypothesis Foam cell formation and atherogenesis malondialdehyde or other TBARS 0: S\_, + LILO macrophage J ca oxLDL O ,f ° ae \ ° scavenger receptor foam cell : plaque formation and ° » atherogenesis
Slide 8: The Oxidative Modification Hypothesis of Atherosclerosis: Does It Hold for Humans?
The Oxidative Modification Hypothesis of Atherosclerosis: Does It Hold for Humans? Joseph L. Witztum* and Daniel Steinberg This review suggests that oxidation of LDL is an important, if not obligatory, event in atherogenesis. The important clinical corollary is that inhibition of oxidation can inhib
Slide 9: Why Haven't the Antioxidant-Based Clinical Trials Been More Effective in Humans?
¢ Why Haven’t the [ Antioxidant-Based] Clinical Trials Been More Effective in Humans? “First, the antioxidants used in these clinical trials may not be sufficiently potent or the doses used may be too low. ... These data suggest that in otherwise healthy people even megadoses of vitamin E are insuff
Slide 10: TBARS predict cardiovascular events in stable coronary artery disease
Journal of the American College of Cardiology. 2004 Nov 16:44(10):1996-2002. Serum levels of thiobarbituric acid reactive substances (TBARS) predict cardiovascular events in patients with stable coronary artery disease: a longitudinal analysis of the PREVENT study. Walter MF, Jacob RF, Jeffers B, Gh
Slide 11: Moderate Antioxidant Supplementation Has No Effect on Biomarkers of Oxidant Damage
Moderate Antioxidant Supplementation* Has No Effect on Biomarkers of Oxidant Damage in Healthy Men with Low Fruit and Vegetable Intakes Robert A. Jacob, Giovanna M. Aiello, Charles B. Stephensen, Jeffrey B. Blumberg,$ Paul E. Milbury, + Lynn M. Wallock+ and Bruce N. Ames+ U.S. Department of Agricult
Slide 12: Effects of Vitamin E on Lipid Peroxidation in Healthy Persons
(lB CLINICAL INVESTIGATION JAMA (Journal of the American Medical Association). 2001; 285: 1178-1182 Effects of Vitamin E on Lipid Peroxidation in Healthy Persons ‘Fina A. Meagher, MD Context Oxidative stress may play a ole in the developmentor exacerbation of many. ‘Orla P. Barry. PhD) common diseas
Slide 13: Self-supplementation with vitamins E & C does not affect TBARS
Self-supplementation with vitamins E & C does not affect TBARS in normal healthy humans 4 R?= 0.802 TBARS (micromolar) 15 1 oe © = supplements, ® (eg, vits E & C) 06 @ = no supplements
Slide 14: No Effect of 600 Grams Fruit and Vegetables Per Day on Oxidative DNA Damage
1016 Vol, 12, 1016-1022, October 2003 Cancer Epidemiology, Biomarkers & Prevention No Effect of 600 Grams Fruit and Vegetables Per Day on Oxidative DNA Damage and Repair in Healthy Nonsmokers! Peter Moller,? Ulla Vogel, Anette Pedersen, Lars O. Dragsted, Brittmarie Sandstrém,} and Steffen Loft Insti
Slide 15: Some compounds induce antioxidant enzymes by producing oxidants
Some compounds induce antioxidant enzymes by producing oxidants Organ damage ke PROOXIDANTS sop malathion, > [ROS]® S carp stanozolol, ete. KL GPx Lipid peroxidation 4 Problem: Even though the antioxidant enzymes are induced, they don’t fully compensate and net injury results. torch
Slide 16: Can the ingredients in Protandim induce the enzymes without toxicity?
Can the ingredients in Protandim® induce the enzymes without paying the price of toxicity and damage? Organ protection sop Protandim — ¢ —— [ROS] [J care [ROS] GPx | Lipid peroxidation The net result would be protection, and oxidative stress would be lowered ric
Slide 17: The induction of human superoxide dismutase and catalase in vivo
Original Contribution The induction of human superoxide dismutase and catalase in vivo: A fundamentally new approach to antioxidant therapy Sally K. Nelson”, Swapan K. Bose *, Gary K. Grunwald ©, Paul Myhill “, Joe M. McCord “4 * webb Waving singe for Cancer. Aging and Ancxidant Research, Univers of
Slide 18: Composition of Protandim
Composition of Protandim® Bacopa moniera extract, 45% bacosides 150 mg Milk Thistle extract, 70-80% Silymarin 225 mg Ashwagandha powder 150 mg Green tea, 98% polyphenols, 45% EGCG 75 mg Turmeric extract, 95% Curcumin 75 mg Total 675 mg Daily dosage for a 70 kg (154 Ibs.) huma
Slide 19: Human Study Design
Human Study Design 29 healthy control subjects were enrolled ranging in age from 20 to 78 years old. The following parameters were measured at days 0, 30, and 120: + TBARS as a measure of lipid peroxidation + Superoxide dismutase (SOD) + Catalase (CAT) + C-reactive protein, an inflammatory marker Ur
Slide 20: 30% Elevation of SOD by Protandim
30% Elevation of SOD by Protandim® 60 45 be rr 100 5 40 L 80 a 5 835 fe} % 30 2 8 25 ‘o 5 20 2 £15 iS 2 5 S10 5 a Qo 30 120 Days on Protandim Average increase in SOD = 30 + 10% after 120 days of Protandim “p< 0.01,n=10 praeiden
Slide 21: 54% Elevation of Catalase by Protandim
54% Elevation of Catalase by Protandim® 140 « z 120 8 ® 100 80 o 0 ‘% Increase in Catalase Change in Catalase (%) 2 3 0 (30 120 Days on Protandim Average increase in catalase = 54 + 15% after 120 days of Protandim *p=0.002,n=10 ric
Slide 22: Inhibition of lipid peroxidation (TBARS) by Protandim
Inhibition of lipid peroxidation (TBARS) by Protandim® 4 R?=0.238 a8 ° OS Before Protandim 3 25 2 18 1 TBARS (micromolar) 0.5 4 After Protandim 30d ao a 20 40 80 80 100 AGE Normal subjects before supplementation with Pratandim showed an age-dependent increase in TBARS. The levels of TBARS dropped an
Slide 23: 40% Average Decrease in TBARS by Protandim
40% Average Decrease in TBARS by Protandim® 4 Plasma TBARS (HM) Before After Protandim Protandim
Slide 24: Achieved outcomes
Achieved outcomes: ee Lipid peroxidation (TBARS) was inhibited ee SOD and CAT were elevated . Possible lowering of C-reactive protein, an inflammatory marker +) Possible sparing of uric acid, a ~ consumable antioxidant ot eae No adverse changes in lipid profile ric