“Development of
Natural Actives for
Spa and Cosmetic
Products”
Chia Wen Chen
Director of Natural Products
Estee Lauder Companies
September 24, 2009
SCC Education Day
R&D Capabilities
Product development assistance
(scale up from a bench top process)
Phytochemical analysis
(to determine marker compound)
Full analytical support
(method development)
Requirements
ISO 9001:2000 certified (cGMP)
Heavy Metal and Pesticide analysis
Safety test for topical application
Sustainable Biomass (wild craft, cultivation)
Certified Organic (NOP, EcoCert, IFOAM)
Challenges
Lot to lot consistency: What are we really getting?
Microbial issues 10,000 cfu for dietary supplement vs 100 cfu
for cosmetic
Heavy metal and Pesticide analysis – due to the pollution in
soil and water
Regulatory (REACH) and Safety (Topical)
Patents- not patent protected
Converting an ingestible for topical use (need to define
biological activity)
Cosmetic Products
Are designed not only to moisturize the skin
and reduce the visibility of lines and
wrinkles…
But also to protect the skin from the
damages caused by environmental
exposure.
Allergen
UV
Pain
Irritant
COX2
IL-1α
AA
PGE2
KC
MN
TNF
IL-1α
PLA2
Nitric oxide (NO)
PLA2
MMP-1
Collagenases
IL-12
TC
PLA2
PL
AA
NO
PMN
FB
NO
TC
Blood flow
CAMs
TC
PG’s
EC
UVB, ozone, smoke
Ozone
PL
A2
UVB
arachadonic acid
O3
O3
O3 O
3
Epidermis
cox2
cigarette smoke
Basement membrane
PGE2
PGE2
neutrophil
collagen
capillary
collagenase
elastase
neutrophil
macrophage
vasodilation
PGE2
Dermis
Control of Inflammatory Reactions
Must take place at the very beginning of the
cascade of reactions.
Inhibition of Phospholipase A2 will result in the
control of the release of Arachidonic Acid, a
major inflammatory mediator.
Additional anti-inflammatories will subsequently
modulate the other steps of the cascade.
Insult (UV exposure, oxidative stress)
Activation of Phospholipase A2
Resveratrol, hops
Poria Cocos
Release of Arachidonic acid
Cyclo-oxygenase
Release of
Prostaglandins
Centaurium
Lipoxygenase
Release of Leukotrienes
Cytokines
(ILI,TNF ∝)
Recruitment of Leukocyte cells
Transport of Leukocytes by blood
Transfer to site of injury (chemotaxis)
Second oxidative burst and Protease (collagenase, elastase, tryptase)
Rosemary, Citri reticulatae peel
Grape seed extract
Premature Aging
The Trend Toward Natural
Your consumers are
watching and learning!
Consumer awareness is at an
all time high.
Consumers want healthy
products - and the industry is
responding.
Natural ingredients and
healthy lifestyle are attractive
to consumers.
Morus spp.
Over 80% of the
world's population
uses plants as their
primary medicines
500,000
plant
species
10,000
substances
per species
What makes plants so efficacious?
multi-taskers
inpired by many
years of folklore
validated by
science
biodegradable
and sustainable
;
High performance ingredients
For high performance skin care
Finding new ingredients
Partnership with consultants and suppliers all
over the world
Strategic alliance with academic research
Rhodiola rosea
OH
O
HO
HO
O
OH
O
O
HO
O
HO
OH
O
HO
OH
OH
rosavin
salidroside
OH
Treatment of skin cells with Rhodiola extract enhanced the ability of
the cells to survive after exposure to an external stress
(UV exposure)
100
% survival
80
60
untreated control
40
Rhodiola rosea
20
0
30
60
90
120
UVB dose (mJ/cm 2)
150
Rhodiola rosea Enhances Rebound from
Stress
number of damaged cells
35
30
25
20
untreated control
15
Rhodiola rosea
10
5
0
50
100
150
200
2
UVB dose (mJ/cm )
After 24 hours the number of damaged cells was
more then
2X lower in the Rhodiola treated skin model
Cellular Damage
(UVB, oxidative damage, inflammatory cytokines)
ATP (energy depletion)
Mitochondrial DNA damage
DNA damage
cancer
(-)Physiological effects
Animation 2
Animation 4
Rhodiola rosea Enhances Rebound from
Stress
Skin model stressed by simulated UV
No treatment
TREATED with
Rhodiola rosea
vs.
Dark brown staining identifies heavily damaged skin cells. The
removal of damage was much faster in Rhodiola treated tissue.
Mimosa tenuiflora
Mimosa bark extract with
Flavonoids, tannins and
saponins.
Anti-collagenase:
EC50 = 80 ug/ml
Collagen synthesis:
31% at 0.0625 µg/ml and
by 67% at 0.25 µg/ml,
Anti-collagenase Activity of Tepescohuite
EC50 = 80ug/ml
% inhibition
100.00
80.00
60.00
40.00
20.00
0.00
0
1.3
4
11.5
34
100
[tepescohuite] µg/ml
310
920
12
Collagen Assay: Tepescohuite
Collagen (µg/ml)
10
8
6
4
2
0
Media
0.015625
0.0625
Dose (µg/ml)
0.25
Stratum
corneum
Langerhans
cell
Mast
cell
elastosis
melanocyte
elastin
collagen
Young skin
Old skin
Loss of collagen & elastin
Photodamaged skin
Degraded collagen, thickened elastin
Young cells
A
vs.
Old cells
B
Fragmented
Fibrils
In vitro cell proliferation as a function of age
20000
Cells/cm2
15000
10000
5000
0
24 Yr
45 Yr
68 Yr
In vitro collagen production as a function of age
Collagen Produced per Cell (ng)
0.14
0.12
0.1
0.08
0.06
0.04
0.02
0
24 Yr
45 Yr
What makes a wrinkle
Constant folding & unfolding of
skin creates a weak point: the microtear.
Physical pressure on cells
activates inflammatory reactions
lead to premature degradation of
collagen by collagenase
Stress induces damages to skin
cells result in further loss of skin
integrity – Wrinkle formation
Wrinkle animation (4) skin folding
development of inflammatory
reaction
Anti-Age: Dermatological Approach
Peels and lasers trigger cell’s natural
defense mechanism.
Skin’s repair process activated by damaged
skin cells.
BENEFIT: improves skin
Inflammatory reaction degrades collagen
& elastin.
SIDE EFFECT: ages skin
Anti-Age: A Natural Approach
Prevent :
control collagenase, elastase and GAGase
degrading enzyme to prevent wrinkle formation.
Restore:
collagen synthesis to accelerate “Healing” of skin
Repair:
help skin cells rebound from stress and prevent
premature aging
Collagen Fragmentation by Collagenase
Healthy skin
Aging/stressed skin
Collagen
Collagenase
Endogenous collagenase inhibitors
ANIMATION 5b collagenase inhibitor
ANIMATION 3 treatment of whole skin; fragments go to
fibroblasts, activate collagen production.
Anti-Age: Wrinkle
Prevent premature destruction of key
dermal components
Treatment:
Anti-collagenase (Sigesbeckia, Tepescohuite)
Anti-elastase (Boswellia, White Birch)
Anti-GAGase (Silymarin, Centaurium)
Collagen synthesis (Tepescohuite)
Result:
Treatment of skin, most damaged by constant
folding / unfolding.
Matricaria recutita flower
Blue chamomile oil with
confirmed:
5-LO inhibitory activity:
EC50 = 50 ug/ml
Anti-chemotaxis:
97% inh. At 0.05%
LPO:
EC50 = 0.003%
German Chamomile
5-Lipoxygenase assay
SCOPE
To analyse potential inhibitors of potato 5-lipoxygenase (5-LO).
PRINCIPLE
5-lipoxygenase converts the substrate linoleic acid (diene 1-4) to a conjugated diene
product (diene 1-3), which can be detected at 234nm. A decrease in the absorbance
signal is indicative for an increased anti-5LO activity.
MATERIAL
Normal lab equipment
96 well half area plates with UV transparent bottom (Costar, 3679)
8 channel multi pipette (20-200µl)
Spectrophotometer
REAGENTS
H2O distilled
Potassium dihydrogen phosphate (Acros, #205925000)
Tween-20 (Janssen Chimica, #2333656)
Dimethylsulfoxide (Acros, #12779)
Potato 5-lipoxygenase (Cayman, #60400)
Linoleic acid 99% (Acros, #215040050)
Nordihydroguaiaretic acid from Larrea divaricata (creosote bush) (approx.
95%)(reference inhibitor - NDGA) (Sigma, N1144)
5-Lipoxygenase Inhibition
IC50 ~50µg/ml
120.00
% Initial 5LO activity
100.00
80.00
60.00
40.00
20.00
0.00
0
10
20
30
40
50
60
70
80
[Blue chamomile oil organic (Hashem Brothers, #4905)] (µg/ml)
90
Chemotaxis Assay: Blue Chamomile Oil Organic
(BRD 4905)
120
100
PMNs/HPF
80
60
40
97% inhibition at 0.05%
20
0
Ctrl
Caff .5%
#4905 .005%
#4905 .05%
LPO (Anti-oxidant)
0.003
0.003
0.002
0.002
0.001
0.001
0.000
30 min
120min
Conclusions
The development of new ingredients from plant
extracts involves many steps which can
ultimately provide great benefits in cosmetics.
Despite the cost involved in such development,
we believe that active ingredients from natural
sources provides the cosmetic industry with a
quasi infinite source of ingredients which we are
aiming at developing in the future.
Conclusion
Opportunity not a Headache
Carefully choose your partner
Know what you are working with
(Standardize extract)