Mangifera indica activates key
metabolic master switch enzymes
The innovation for well-aging
Dr. S. Buchwald-Werner
6. May 2015
Vitafoods Europe Conference – Healthy Aging Session
Consumer demand for healthy-aging products
World´s population is aging rapidly and life expectancy is increasing
Aging consumers are the physically and mentally fittest in history
Today all age-groups take personal responsibility for
their health and are interested in products to reduce
the signs of aging and the risk of metabolic diseases
There is a 7 years gab between
Life expectancy & “healthy” life expectancy
In 2050 the proportion of people over 60 years is expected
to increase from 605 million to 2 billion until 2050
There is a market for well-aging products
Jane Fonda, 75 years, 2014 http://www.welt.de/lifestyle/article113599606/Jane-Fonda-fast-zu-perfekt-um-75-zu-sein.html Foto: Invision
WHO 2012: Interesting facts about aging. http://www.who.int/ageing/about/facts/en/#; 18.08.2014O
Euromonitor International Life expectancy 2013
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Agenda
1. What happens during aging?
2. Emerging biological targets to develop ingredients for well-aging
3. Mangifera indica – the natural well-aging ingredient for application
in food supplements and foods; Ingredient introduction and
scientific results for

Key metabolic master switch enzyme: SIRT 1

Key metabolic master switch enzyme: AMPK

Mitochondria and aging
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1. What happens during aging?
Biological aging is commonly defined as
the successive accumulation of adverse
effects in an organism with increasing age
Our cells lose power because
less power plants (Mitochondria)
are produced and the remaining
are less efficient
Our cells lose direction
because mistakes in DNA
replication by oxidative stress
and inflammation are adding
up to first disturbances in our
metabolism
Enzymes and metabolic master switches
decline, leading to an increase of blood lipids
and body fat mass in combination with
reduced muscle mass as well as elevated
blood glucose and insulin resistance
Jane Fonda, 75 years, 2014 http://www.welt.de/lifestyle/article113599606/Jane-Fonda-fast-zu-perfekt-um-75-zu-sein.html Foto: Invision
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Agenda
1. What happens during aging?
2. Emerging biological targets to develop ingredients for well-aging
3. Mangifera indica – the natural well-aging ingredient for application
in food supplements and foods; Ingredient introduction and
scientific results for

Key metabolic master switch enzyme: SIRT 1

Key metabolic master switch enzyme: AMPK

Mitochondria and aging
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2. Emerging biological targets for well-aging
Master switches, e.g. SIRT1, AMPK:
Evolutionary conserved metabolic sensors
that steer energy homeostasis by
„instructing“ the body to burn or not to burn
energy and consequently to maintain
essential body functions
Mitochondria:
working organelles
that keep the cell
full of energy
The function and survival of all organisms is dependent on the
dynamic control of energy metabolism, when energy demand
matches energy supply
These master switches have homologs in all eukaryote;
evolutionarily conserved - all the way from yeast to humans
Effective mitochondrial biogenesis and turnover is critical for
the maintenance of energy production, the prevention of
oxidative stress and well-aging
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Agenda
1. What happens during aging?
2. Emerging biological targets to develop ingredients for well-aging
3. Mangifera indica – the natural well-aging ingredient for application
in food supplements and foods; Ingredient introduction and
scientific results for

Key metabolic master switch enzyme: SIRT 1

Key metabolic master switch enzyme: AMPK

Mitochondria biogenesis
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3. Mangifera indica for well-aging
100% Mango fruit powder (Mangifera indica L.)
Mangos are harvested at a special degree of ripeness
from selected farmers to obtain high concentrations of secondary
plant compounds which are important for the health benefits
Several scientific studies have been carried out using the proprietary
ingredient to investigate the effects of well-aging
Nutritional benefits (fiber, vitamins and minerals)
A high quality, sustainable natural ingredient
Approved for use in dietary supplements & foods
Careless™ is the first scientifically proven and IP protected ingredient
that helps you care less about aging
while delivering the great taste and pleasure of exotic mango fruits
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3. Key metabolic master switch enzyme: SIRT 1
Sirtuin structure (MIT)
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3. Key metabolic master switch enzyme: SIRT 1
Reduction
of body fat
Increased
muscle mass
Balanced
blood sugar
& insulin
NAD+
Nicotinamid +
ADP-ribose
SIRT1
Transcription/ Co-factors:
e.g. PGC-1a
FOXO
LXR
PPARg
p53
Beta-cell
protection
Deacetylated
targets
Cholesterol
homeostasis
Endothelial
function
Boutant et al. Molecular Metabolism 2013
Li X Aca Biochim Biophys Sin 2013
Sirtuin structure (MIT)
Hubbard ad Sinclair, Trends in Pharmacological Sciences (2014) 35:
DNA repair
Cell protection
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3. Mangifera indica for well-aging
Activation of SIRT 1 using human recombinant enzyme
Enzyme deacetylation activity was compared,
before and after treatment with a water extract
based sample preparation of
4 different batches of Mangifera indica fruit powder
and resveratrol as positive control.
Optimal sample concentration of 500µg/ml was determined
in a pre-test
Two different test systems were used:
1. Fluorescence assay
SIRT1 active recombinant human
protein expressed in E.coli
2. Luminescence assay
SIRT1 active recombinant human
protein expressed in Sf9 cells
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3. Mangifera indica for well-aging
Activation of SIRT 1 using human recombinant enzyme - Results
Fluorescence assay
All Mangifera fruit powder batches
activated SIRT1 by > 30%
Resveratrol ~50%
Luminescence assay
All Mangifera fruit powder batches
activated SIRT1 by > 40%
Resveratrol ~95%
Results show that Mangifera indica fruit powder is able to activate SIRT1
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3. Key metabolic master switch enzyme: AMPK
Steinberg GR, Kemp BE, AMPK in Health and Disease, Physiological Reviews (2009)89 : 3, 1025-1078
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3. Key metabolic master switch enzyme: AMPK
Improved
hepatic fatty
acid
oxidation
Cholesterol
Triglyceride
homeostasis
P
AMP: ATP
Improved
muscle
fatty acid
oxidation
AMPK
AMPK
Kinases / Ca2+
Transcription/ Co-factors, e.g.
ACC
COX
PGC-1a
GLUT1,4
mTOR
HMG-CoA
Improved
muscle
glucose
uptake
Balanced
blood sugar
& insulin
Steinberg GR, Kemp BE, AMPK in Health and Disease, Physiological Reviews (2009)89 : 3, 1025-1078
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3. Mangifera indica for well-aging
Activation of – AMPK in human hepatic progenitor cell line (HepaRG)
Enzyme activity was compared (phosphorylation)
before and after treatment with a water extract
based sample preparation of Mangifera indica fruit powder
and resveratrol and metformin as positive control
Cytotoxicity test was carried out
Dose-dependency was tested (30µg/ml, 100µg/ml, 300µg/ml)
Activity of 3 different batches was tested
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3. Mangifera indica for well-aging
Activation of – AMPK in human hepatic progenitor cell line (HepaRG)
Results:
Mangifera fruit powder activated
AMPK in a dose dependent manner,
up to 100 %
Resveratrol and Metformin ~100%
All Mangifera fruit powder batches
activated AMPK by > 50%
Resveratrol ~82%
Metformin ~130%
Results show that Mangifera indica fruit powder is able to activate AMPK
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3. Mitochondria and aging
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3. Mitochondria and aging
Mitochondria - processes
Heat
Production
Thermogenesis
Energy
ATP
Production
Calcium
storage
ROS
homeostasis
Apoptosis
mt DNA
cellular
Signaling
Hormones
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3. Mangifera indica for well-aging
Mitochondrial biogenesis in liver and muscle cell lines
Capacity to activate mitochondrial biogenesis
before and after treatment with a water extract
based sample preparation of Mangifera indica fruit powder
Cytotoxicity test was carried out
In murine muscle cell line (C2C12)
dose-dependency was tested (200µg/ml, 400µg/ml);
resveratrol as positive control
In human hepatic cell line (HepG2)
dose-dependency was tested (100µg/ml, 200µg/ml, 400µg/ml)
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3. Mangifera indica for well-aging
Mitochondrial biogenesis in liver and muscle cell lines - Results
In murine muscle cell line (C2C12)
Mangifera fruit powder enabled
mitochondrial biogenesis > 7 %
Resveratrol up to 17%
In human hepatic cell line (HepG2)
Mangifera fruit powder enabled
mitochondrial biogenesis
dose dependent > 15 %
Results show that Mangifera indica fruit powder enabled mitochondrial biogenesis
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3. Mangifera indica for well-aging
Summary
Reduction of
body fat
Increased
muscle mass
Well-aging
Balanced
blood sugar
Oxidative
stress
resistance
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3. Mangifera indica for well-aging
R&D Outlook
1. Phytochemical investigation
2. In vitro study to investigate eNOS activation
3. In vivo animal study to investigate weight management - ongoing
Body weight and Body fat
Gene expression to confirm mode of action (SIRT, AMPK)
4. Human study – Well-aging - ongoing
Skin microcirculation acute study
Vascular function acute study
5. Human study – Weight management (to be defined based on animal study results)
6. Human study – well-aging (to be defined based on 1st human study results)
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Vital Solutions - Company introduction
Vital Solutions develops innovative condition specific natural ingredients for
use in foods and dietary supplements, which serve consumer demands for
health products and help to reduce health care costs
Based on a market-oriented dynamic strong R&D platform, we focus on the
development of natural ingredients which improve quality of human life by
feeling good and aging well. Our research is focusing on:
 Anti-aging
 Brain health
 Digestive health & metabolism
Over the past 5 years we have created an innovative product portfolio
and a pipeline, capable of delivering sustainable sales growth.
 Market introduction of 4 innovative health care ingredients
 Set up of an innovative product pipeline of 4 ingredients
 Global distribution network
Latest innovation: Ingredient for healthy-aging
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Thank you for your attention!
Meet us at Vitafoods
Booth G51
Vital Solutions GmbH
Dr. Sybille Buchwald-Werner
Hausinger Straße 6
D-40764 Langenfeld
Phone: +49 (0) 2173 10 98 202
E-Mail: [email protected]
All information given in this presentation reflects our current knowledge and interpretation of relevant legislations and legal
practice. Vital Solutions expressly disclaims any responsibility for the suitability of this information for products and for any specific
or particular purposes intended by the user. Our suggestions do not release Vital Solutions´ customers from evaluating the
individual products as to their legal compliance. Vital Solutions does not assume any liability or risk involved in the use or
marketing of its products as the conditions of use are beyond its control. The user of the products is solely responsible for
compliance with all laws and regulations applying to the use of the products.
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