Modification of the NAD+/NADH ratio to
mimic Calorie Restriction
An Update for 2014
Alan Cash
Terra Biological LLC
 Part
1 – Review of Mechanism of Action:
Using Oxaloacetate to increase the
NAD+/NADH Ratio
 Part
2 - Current Applications of Oxaloacetate
as a Nutritional Supplement– Specific
Information to support your practice
 Part
3- Clinical Trial Update for Possible
Future applications
 Part
4- Patient Profiles that may benefit
Review of
Mechanism of Action
 2009
Aging CELL
“Oxaloacetate Supplementation Increases
Lifespan in C. elegans through an AMPK/
FOXO-dependent pathway”
 2009
Open Longevity Science
“Oxaloacetic Acid Supplementation Mimics
Calorie Restriction”
 2010
Anti-Aging Therapeutics
Chapter 6 “Modification of NAD+/NADH”
Benefits:
 Increases in average and maximal lifespan—
up to a 40% increase is seen in mammals.
 Decreases in cancer incidence—up to a 55%
decrease.
 Decreases in neurodegenerative diseases
such as Alzheimer’s and Parkinson’s disease.
 Complete protection against type II diabetes.
 Reduction in cardiovascular risk and, in
particular, atherosclerosis.
 Reduction in inflammatory diseases, such as
auto-immune type diseases.
 NAD+/NADH
h in calorie restriction, Lifespan h
Lin 2003, Easlon 2008, Edwards 2013, Lee 2012,
Braidy 2011
 NAD+
Salvage h human cell lifespan h
Van der Veer 2007
 NAD+
precursors h lifespan h
Belenky 2007
 AMPK
h Lifespan h
NAD+/NADH h AMPK h
Greer 2007, Rafaeloff-Phail 2004
 Mitochondrial
Yang 2007
NAD+ h Cell survival h
 NAD+/NADH
i Sarcopenia h
Pugh, 2013
 NAD+/NADH
h mitochondrial density and
Energy h
Chuang 2013
 NAD+
i mitochondrial OXPHOS i
Gomes 2013
 NAD+/NADH
h breast cancer metastasis i
Santidrian 2013
 How



to increase the NAD+/NADH Ratio?
Calorie Restriction (via glucose starvation)
Prolonged Exercise (via Gluconeogenesis)
Supplementation with oxaloacetic acid
Delta G = -29.7
Malate
Dehydrogenase
Oxaloacetate
Malate
NAD+/NADH Ratio
NADH
NAD+
Oxaloacetate
Supplementation
increases
NAD+/NADH
ratio by 900%
Krebs, 1968
Ascorbic
Acid
Oxaloacetate
Cell Membrane
AMPK
Activation
Oxaloacetate
NADH
DAF-16
Transcription
Factor
Oxaloacetate
Malate
Dehydrogenase
NADH
Malate
Dehydrogenase
?
DG
-29.7
DG
-29.7
?
Life
Extension
NAD+
Malate
Cytosol
Malate
Inner Mitochondrial
Membrane
Mitochondria
NAD+
98% Positive Directional
Overlap for genes
Changed in common.
OAA
Gene
Symbol
Foxa1
Foxa3
Foxq1
Foxq1
Gene Title
forkhead
forkhead
forkhead
forkhead
Affymatrix
Gene
Number
2891
13370
6994
30006
Change in
Gene
Expression
Calorie
Restricted to
Control
30% Increase
100% Increase
110% Increase
190% Increase
Change in
Gene
Expression
benaGene to
Control Gene function
40% Increase
regulation of transcription,
DNA-dependent // inferred from
electronic annotation
70% Increase
cell glucose homeostasis //
inferred from mutant phenotype
// regulation of transcription,
DNA-dependent // inferred from
mutant phenotype /// cellular
response to starvation // inferred
from mutant phenotype
210%
Increase
regulation of transcription,
DNA-dependent // inferred from
electronic annotation
220%
Increase
regulation of transcription,
DNA-dependent // inferred from
electronic annotation
Applications of Oxaloacetate
as a Nutritional Supplement
h
The Krebs Cycle
(Citric Acid Cycle)
Percent Survival
100
OAA Effect on C. Elegans
75
8mM
OAA WT
50
WT
25
16mM
OAA WT
0
Days Alive
OAA effect on Mice
25% increase
Mouse Data:
Steve Spindler
UC Riverside
Increase in Lifespan
140%
130%
120%
110%
100%
90%
80%
70%
Worms
Flys
Mice
Clinical Trial– Diabetic Patients
Decrease in Fasting Glucose Levels
70.00%
60.00%
50.00%
40.00%
30.00%
20.00%
10.00%
0.00%
1
2
3
4
5
6
7
8
9
10
11
12
13
14
Average decrease of 24%
15
16
17
18
19
20
21
Fasting Blood Glucose Measurement
Non-Diabetic Patient
100
95
mg/dL
90
85
80
begin OAA here
75
Before OAA
2-3 Week Transition Period
After OAA
70
1
3
5
7
9
11
13
15
17
19
21
23
25
27
29
31
33
35
37
39
Laboratory Research
Translates into
Commercial Product
www.benaGene.com
 73
year-old European Woman
 Currently on Anti-Diabetic medicines
 Diaprel MR/ (80 mg)- 2 per day – Extended
release Gliclazide (80 mg), a once per day
diabetic drug
 Merckformin (1,000 mg)- 1 per day(Metformin, Glucophage) – used for Type 2
diabetes

Avandamet- 1 per day- a combination of
metformin and rosiglitazone used for diabetes
Glucose Levels
18
16
14
Fasting
Breakfast
bed
Linear (Fasting)
Linear (Breakfast)
Linear (bed)
mmol/L
12
10
8
6
4
2
0
0
20
40
60
80
 Fasting
glucose levels dropped 23%.
 Glucose
levels after a meal dropped 34.5%,
indicating a major improvement in glucose
management and glucose tolerance.
 Metformin
use reduced by 50% during the
study.
 Results
are statistically significant.
20-30% Decrease in Genes that Create & Store Fat
29% Reduction in Fat Tissue Mass with OAA Supplementation
Reduced Weight Rebound Due to Lower Gene Expression
After Diet
Commercial Product
 OAA
protects mitochondrial DNA in the brain
Yamamoto 2003
 retinal
pigmented epithelium (RPE), damaged
in age-related macular degeneration (AMD)
are protected by zinc and OAA
Wood 2003
 pancreatic
islet cells and neurons are
protected by OAA

Chang 2003, Berry 2006
 OAA

is a powerful anti-oxidant
Desagher 1997, O’Donnell-Tormey 1987
 10%
increase in Muscle Endurance with OAA
 AMPK activation as Precursor to Mitochondrial
Biosynthesis
*
240
235
230
225
220
215
210
205
200
195
Control
OAA
Imagine what a 10%
Increase in Endurance
Does for competition.
Reduction in Essential Tremor
www.TremorStop.com
Clinical Trial Updates for
Oxaloacetate as a Potential
Drug for Disease Treatment
 Potential






Support for:
Cancer
Alzheimer’s
Parkinson’s
Stroke
Epilepsy
Diabetes
 Animal
Data looks good, but still early in the
research effort.
 These statements have not been evaluated
by the FDA. Oxaloacetate is not intended to
diagnose, treat, cure or prevent any disease.
 Calorie

Restriction deceases cancer risk
Currently one of the most effective broad-based
methods to reduce cancer risk
 High
NAD+/NADH ratio decreases cancer
Metastasis Santidrian 2013
 Oxaloacetic Acid Supplementation prevents
Human Lung Cancer cells from reproducing




In vitro results
Does not affect normal cells
Prevents replication of cancer cells by increasing
intercellular debris, but does not kill the cells.
Cancer cells did not reproduce after OAA solution
removed for six weeks.
Farah 2007
OAA Reduces Glioblastoma Tumor Growth by 50%
Rubin, Inv. New Drugs
2012
21 day tumor growth in nude mice. Ruban, Investigative New Drugs 2012
Temozolomide
And
Oxaloacetate
No tumor cells found
In Remaining Mice.
Rubin, Invest. New Drugs 2012
After Review, US FDA
Designates
Oxaloacetate
As an “Orphan Drug”
for the
Treatment of Gliomas.
Designation (12-3704)
Medical Foods are a
unique category
- Allow immediate
implementation
- Food Additives or GRAS
only
- Used under Physician
Supervision
- Must have a Scientific
Basis
 Case
studies indicate that Gliaxal, either alone or
in combination with other therapies, stopped
tumor growth in 88% of the patients (N = 17)
44
F
Anaplastic Astrocytoma
Surgery, Radiation and
Chemotherapy
None
7 months
No Growth
44
M
Glioblastoma
Surgery, Radiation and
Chemotherapy
Temodar
6 months
No Growth
44
M
Glioblastoma
Surgery, Radiation and
Chemotherapy
Everolimus, XL184
5 months
Tumor Reduction
62
M
Glioblastoma
Surgery, Radiation and
Chemotherapy
None
4 months
Progressive
34
F
Anaplastic Astrocytoma
Surgery, Radiation and
Chemotherapy
Everolimus, XL184
4 months
Tumor Reduction
26
M
Low-grade Oligodendroglioma
None
4 months
No Growth
Surgery
62
M
Glioblastoma
Surgery, radiation and
Chemotherapy
69
F
Glioblastoma
Surgery, radiation and
Chemotherapy
None
2 months
No Growth
36
M
Low-Grade Oligodendroglioma
Surgery, Radiation and
Chemotherapy
Temodar
2 months
No Growth
Ketogenic Diet
4 months
9 months
No Growth
24
M
Anaplastic Astrocytoma
44
M
Glioblastoma
Surgery, Radiation and
Chemotherapy
Temodar
2 months
No Growth
50
M
Glioblastoma
Surgery, Radiation and
Chemotherapy
Nilotinib
1 month
No Growth
51
M
Surgery, Radiation and
Chemotherapy
Temodar
1 months
No Growth
Low-Grade Oligodendroglioma
Surgery
Temodar
Tumor Reduction

"My son has an astrocytoma. With a ketogenic diet and
significant nutritional support including Gliaxal Medical
Food, the last two MRI's have showed no change and
some small amount of shrinkage. The Oncologist says
it is very slow growing tumor and is not gung-ho on any
aggressive action. He said to keep up what we are
doing and follow-up MRI in 4 months. It's been tough
trying to keep a 24-year old healthy kid on the program,
but he is doing pretty well. We plan to continue with
Gliaxal. We have used 2 to 10 capsules three times per
day with no side effects. THANK YOU.”
Dr. Loren Stockton
 Starting
a Phase 1 clinical trial in Pediatric
Brain Cancer this summer at Rady Children’s
Hospital, San Diego
 Continuing
case studies at UCSD and George
Washington University
Substantial Improvement in Quality of Life
Mobility
Activities of Daily Living
Emotional Well Being
Stigma
Before
After
Social Support
Cognitive Impairment
Patient QOL Impact
Score Improved From
79 to 8
Communication
Bodily Discomfort
0
5
10
15
20
25
30
35
 Effective
in 54% of patients surveyed (N=13)
 In
the Effective Group, the average
improvement in Quality of Life score was 63%
(P < 0.05)
 Range
of improvement varied between 20%
and 90%
 Currently
in Phase II Clinical Trial, See
http://clinicaltrials.gov/ct2/show/NCT01741701
(Pistel, 2012)
(Pistel, 2012)
(Pistel, 2012)
*
(Pistel, 2012)
*
Alzheimer Model Mice show memory improvements
50% Increase in Short-Term Memory
(Pistel, 2013)
A
Clinical Phase 1 trial is beginning with
oxaloacetate supplementation and
Alzheimer’s disease at the University of
Kansas.
Patient Profiles that may
receive benefit from
oxaloacetate
 As




Anti-Aging – General Population
Helps to maintain proper glucose function
Improves Mitochondrial Function and Density
Weight Maintenance






Reduces the “Rebound Effect” After Dieting
Increases Endurance & Fights Fatigue


a Nutritional Supplement
Elderly Patients
Fatigued Patients
Athletes
Mood swings within PMS
Constipation Reduction
Glutamate Reduction
 As






an Experimental Drug Clinical Trial
Parkinson’s Disease
Glioblastoma
Alzheimer’s Disease
2-hydroxyglutaric aciduria
Mitochondrial disease
Diabetes
 Necessary




for Clinical Trial
IRB Approval
Data sent to ClinicalTrials.gov
Patient Approval Form
30-day FDA notification prior to the trial
Alan Cash
Terra Biological LLC
San Diego, CA 92130
858-947-5722
acash@benaGene.org
Web:
www.benaGene.com
www.Gliaxal.com