Download A Statewide Systems Biology Approach

Driving the Future of Nutraceuticals
The Institute for
Nutraceutical Research at
Clemson University
Driving the Future of Nutraceuticals
A Statewide Systems Biology Approach
Medicinal Plants with Anti-inflammatory
and Anti-oxidant Properties
Culinary Herbs
Vegetables
Traditional
Medicinal Plants
Turmeric (curcumin)
Cilantro/Coriander
Leafy & other
crucifers
Muscadine
Yarrow
(broccoli, collards,
kale etc.)
Neem
Fennel
Watermelon
Noni
Primrose
Tomato
Bitter gourd
Tarragon
Peppers
Mahogany
Garlic
Black cohosh
Bloodroot
Suppression of NF-B and Inflammation by spice
polyphenols, including the Indian spice turmeric (currie)
Fennel, (anise,
coriander)
Red Chilli
(capsaicin)
Garlic (diallyl
sulfide; ajoene,
allicin)
Turmeric
(curcumin)
Cloves (Eugenol
& isoeugenol)
NF-B blockers
from spices
Ginger (6gingerol)
Pomegranate
(Ellagic acid)
Basil &
rosemary
(Ursolic
acid)
Aggarwal, BB & S. Shishodia. Ann NY Acad Sci. 1030:434-441, 2004
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Meeting Today’s Challenges, Providing Tomorrow’s Solutions
“Cellular and Animal Model Screening of
Potential Performance-Enhancing
Phytonutrients”
DoD Combat Ration Research & Development
“Basis for & extent which food constituents and dietary
supplements:
-Delay fatigue
-Extend physical strength & endurance or
-Heighten alertness or enhance cognitive abilities of
soldiers engaged in physically or mentally
demanding tasks”
Driving the Future of Nutraceuticals
High Throughput Screening of
Phytonutrients
Level I- Immune Cell Markers
Pro-inflammatory and Anti-inflammatory
cytokine release
Antioxidant properties
NF kappa B inhibition
Prostaglandin inhibition
Level II- Animal Models
Physical stamina and recovery from fatigue
Mental alertness and cognitive functions
Resistance to viral infection
Examples of Purified Polyphenols Tested
Driving the Future of Nutraceuticals
Example of Mixed Polyphenols in
Muscadine
Skin Only
Muscadine Polyphenols
(mg/100g FW)
Seeds
Skins
Leaves
Pulp
Antioxidant Capacity
281**
13
236
3
Total Phenolics
2180**
375
352
24
Gallic acid
7
Catechin
560*
Epicatechin
1300*
9
Ellagic acid
17*
67
Myricetin
8
158*
Quercetin
2
10
Kaempferol
0.6
9
Trans-resveratrol
0.1
Anthocyanins
132*
Anthocyanidins
Compounds
mg/g of muscadine extract
Malvidin
Delphinidin
Cyanidin
10.39
5.57
3.62
12
Promising Anti-inflamatory and
Anti-oxidative Nutraceuticals
Agent
INF áINF* NF-kB Recovery**
Muscadine - ++++
++++
Curcumin
+++
+++
Quercetin
+++
+++
Caffeine
++
Echinacea +++
Astragalus ++++ * Inhibition of cytokines and prostaglandins
** Recovery from fatigue
RESPIRATORY VIRUSES OF MILITARY IMPORTANCE
Adenovirus
Influenza Virus
Rhinovirus
Proposed Mechanism of the Cytokine Storm Evoked by
Respiratory Viruses
Osterholm, M. T.
N Engl J Med 2005;352:1839-1842
Quercetin Enhances Resistance to
Influenza Following Exercise Stress
120
Ex-Placebo
Ex-Quercetin
Con-Placebo
Con-Quercetin
100
% HEALTHY
80
]*
]*
60
40
Davis, Murphy &
Gangemi, Am. J.
Physiol. in press
*
20
0
0
5
10
15
DAY POST-INFECTION
20
25
Key Elements in Rhinovirus
Pathogenesis
• Virus replication in nasal mucosal cells
(temp. restricted, 33- 350 C)
• Replication induces oxidative stress in
respiratory epithelium
• Activation of NF Kappa-B> amplifies immune
response genes
• Induction of IL-8
• Recruitment of PMNs/degranulation
Quercetin Suppression of Rhinovirus-Induced
IL-8 Elaboration in Respiratory Cells
5000
High MOI, early collection
Low MOI, late collection
3000
2000
1000
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IL-8 concentration
(pg/ml)
4000
Next Steps
• Confirm antiviral activity in experimental
adenovirus infections
• Examine clinical effects in natural cold model
(adenovirus and rhinovirus)
• Evaluate alternative delivery models (i.e.
buccal waffers) for improved bioavailability
• Genetically modify plants (e.g. muscadine)
to produce higher concentrations of
secondary metabolites
Brain/Muscle Food - Best Bets
• Carbohydrates – Good bet to delay mental and
physical fatigue
• Caffeine – Good bet to delay mental
fatigue
• Optimal doses vary widely and side effects
can be serious in high doses, especially if
combined with other stimulants
• Quercetin – Good evidence of a benefit
on mental and physical fatigue
• Curcumin – Some evidence of faster performance
recovery following muscle damage
• Muscadine Extract – Rising fast
Cellular Oxidant Stress in
Response to
Rhinovirus Challenge
Cell control
Rhinovirus
Challenged
Carbonyl stain
DCFDA stain
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Geometric Mean Virus Titer
TCID50/ml
Effect of Quercetin on Rhinovirus
Replication in Respiratory Cells
High MOI, early collection
Low MOI, late collection
3
2
1
0
Curcumin from Q-Chews
Quercetin from Q-Chews
Muscadine
Curcumin from Q-Chews
Quercetin from Q-Chews
26
Curcumin from Q-Chews
Quercetin from Q-Chews
Muscadine
Curcumin from Q-Chews
Quercetin from Q-Chews
28
Q-Chews
Percent Inhibition of DNA Damage (%)
Percent Inhibition of DNA Damage (%)
100
80
60
IC50
40
20
0
-3
-2
-1
0
1
log Concentration of Myr (M)
IC50 = 2.0 ± 1 µM
2
3
100
80
60
IC50
40
20
0
-3
-2
-1
0
1
2
log Concentration of Q (M)
IC50 = 10.8 ± 1 µM
Physiological concentrations of polyphenols commonly reach 1-10 M
Yamamoto, T.; et al. J. Pharm. Exp. Ther. 2003, 307(1), 230-236.
Scalbert, A. and Williamson, G. J. Nutr. 2000, 130, 2073S-2085S.
Driving the Future of Nutraceuticals
3
• Quercetin improves cycling
time trial performance in
humans
• Green tea supplementation
reduced muscle lipid
peroxidation in aerobically
exercised rats
Peake, J.M.; et al. J. Nutr. Biochem. 2007, 18,
357-371; Urso, M.L.; et al. Toxicology 2003,
189, 41-54; Leeuwenburgh, C.; et al. Curr.
Med. Chem. 2001, 8, 829-838. MacRae, H.
S.-H.; et al. Int. J. Sport Nutr. Exer. Metab.
2006, 16(4), 405-419. Chai, Y.-M.; et al. J.
Food Sci. Nutr. 2003, 8(4), 377-382.
Driving the Future of Nutraceuticals
Food factors affecting immune status
Nutrient
components
Amino acids
Glutamine
Arginine
BCAAs
Proline
Lipids ω-3
PUFAs
Vitamins A,
B, C, D, E
Immune
status
Glutathion
e
Non-nutrient
components
Minerals Fe,
Zn, Se
Flavonoids
isoflavones
genistein
• 100% DNA damage inhibition is observed for all polyphenols
at concentrations of 50 – 500 µM with Fe2+/H2O2
• Gallate compounds were more potent than their catecholate
analogs in every instance
• IC50 values correlated to pKa values of the most acidic
phenolic hydrogen for catecholate compounds
• Both iron-binding and lipophilicity are important factors: iron
binding determines antioxidant activity, while lipophilicity
determines bioavailability
• Have also developed protocols for testing antioxidant activity
against DNA damage from both Cu+/H2O2 and peroxynitrite
Driving the Future of Nutraceuticals
HOST DEFENCE SYSTEM
Nonadaptive
Adaptive
Innate
lymphocytes
cytokines
Surface Alternative Macrophages
epithelia pathway of (monocytes)
complement
activation
Dendritic
cells in skin
and
lymphoid
tissue
B
T
Plasma
activated
cells
antibody
subsets
producing
CD4
CD8
interferons NK cells
(natural
killer cells)
and certain
B + T cells
Chemopreventive activities of antioxidants
(Food Ingredients to Stimulate Protectve athways)
(Adapted from Brash and Havre, PNAS 2002; 99,13969)
Dietary Antioxidant Activities
Redox
Regulation
Radical
Scavenging
Redox
Regulation
Reduced DNA, Protein &
Membrane Damage
Proline-linked
Pentose-phosphate
P53
Pathway
Antimicrobial
Activities
Cox-2
Antiinflammatory
Pathways
DNA
Repair
Prevention
Dietary Health Management
Transformation-specific
Apoptosis
Restitutio
n
“It isn’t what you
eat that can kill
you, and it isn’t
just your DNA
that can save
you- it’s how
they interact”
Jose Ordovas, 2006
Research Partnership For Preventing Cancer With
Botanicals
“SC Grown”
Fruits and Vegetables
“SC Grown”
Culinary Herbs
Traditional Medicinal
Plants
MW 1
2
3
4
5
6
7
8
9
OH
10 11 12 13 14 15 16 17 18
OH
[Q]
HO
O
OH
OH
O
Quercetin (Q)
Control Lanes: MW = 1 kb DNA ladder; 1 = plasmid DNA; 2 = DNA, 50
M H2O2; 3 = DNA, 500 M Q, 50 M H2O2; 4 = DNA, 2 M Fe2+, 50
M H2O2
Experimental Lanes: 5-18 = DNA, 2 M Fe2+, 50 M H2O2, and
increasing Q concentration (0.001, 0.002, 0.02, 0.05, 0.1, 0.2, 2, 4, 10,
50, 100, 200, and 500 M, respectively)
Driving the Future of Nutraceuticals
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1500
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IL-8 concentration
(pg/ml)
Effect of Quercetin on Rhinovirus-Induced
IL-8 Elaboration in NHBE Cells
High MOI, early collection
Low MOI, late collection
1250
1000
750
500
250
0
NADH
Fe2+
H2O2
NAD+
Fe3+
OH
•
Iron-mediated DNA damage is the primary
cause of cell death under oxidative stress
•
Inhibiting this damage may prevent tissue
damage from heart attack or stroke,
neurodegenerative diseases (Alzheimer’s,
Parkinson’s), cancer, aging, fatigue and
impaired recovery from exercise
Henle, E. S.; et al. J. Biol. Chem.
1999, 274, 962-971; Orrenius, S.; et
al. Annu. Rev. Pharmacol. Toxicol.
2007, 47, 143-183; Park, S.; Imlay,
J. J. Bacteriol. 2003, 185, 19421950. Valko, M.; et al. Chem.-Biol.
Interact. 2006, 160, 1-40.
Driving the Future of Nutraceuticals