PROGELINE™

Modulates progerin, a marker of senescence

ANTI-AGING/ ANTI-WRINKLING/ CELL TURNOVER

Progeline™

•The anti-aging market

•The concept of senescence & progerin

•Efficacy summary

•Clinical efficacy

•Tolerance studies

•Applications

•Conclusion

2

The Anti-aging market

Among the global anti-aging market, the appearance segment is expected to generate

$105B in 2013, for an annual growth rate of 10,4% for the past years.

Europe was the largest anti-aging skincare market in 2010, with sales of €2.4B

The US was close behind, with mass market sales alone of $804m.

The number 55-64 years old will grow 31% in the US, while the 65-74 year old group will increase by 31%

A similar situation will emerge in Western Europe by 2013.

Anti-aging launches:

Source: Mintel GNPD Beauty Innovation, Jan 2007 – Dec 2010, anti ageing face/neck and eye care, France, UK, USA, Japan, China

The state of anti-aging market…

Dior has added stem cell + growth factor terminology to the product’s

original detox concept

PaloVia Skin Renewing Laser

FDA-cleared device and gel claimed to reverse the signs of aging

Esthe Pro Labo Outer Ageing Care Program

New skincare range with moisturising horse-derived placenta

• Growth Factors • Stem cell activators • Sirtuins activators

• Telomerase activators & DNA repair • Neurocosmetics • Anti-aging device & laser technology

These days the fight against aging is addressed through different mechanisms of action:

In 2012, the fight is no longer only about signs of aging, but in the fight to delay senescence processes…

FOCUS

ON

PROGERIN

5

6

Where to find a new senescence marker?

Progerin is a senescence marker discovered through the understanding of Progeria (an acceralated aging condition in children)

Senesecence is characterized by a build-up

of a toxic protein progerin, which is critical

for the cellular activity (nucleus structure and

DNA replication).

The accumulation of progerin causes nuclear

defects and DNA damages, thus contributing to

the appearance of signs of aging. Cell nucleus of a

9 years old Cell nucleus of a

96 years old

Progerin is present in fibroblasts and keratinocytes.

It accumulates with time with higher levels later in life.

Progerin expression is now used as biomarker of cellular aging.

Progeline™ Rethinking skin aging by modulating progerin; the senescence marker

Progeline™, a 3 amino acids peptide biomimetic of Elafin, an enzyme inhibitor involved in extracellular matrix integrity.

Progeline™ decreases progerin synthesis, a new aging biomarker involved in skin cellular senescence.

By acting directly on the senescence process, Progeline™ clinically improves the signs associated with skin maturation: sagging, slacking and wrinkles.

INCI name: Glycerine (and) Water (and) Dextran (and) F3Acetyl Tripeptide-2

Protocol

Mature human dermis fibroblasts were incubated for 96 hours with different ingredients.

The progerin was quantified in cell lysates using an ELISA methodology.

Results are given in pg of progerin per μg of proteins

Progeline™ decreases significantly the production of progerin, attesting its potential as anti-aging ingredient!

Reduction of progerin production

Young and healthy skin…

Dermis level:

↑ collagens = gives the skin its strenght

↑ elastin = gives the skin its elasticity

Healthy and radiant looking skin

Epidermis-DEJ level:

Thick Epidermis = proper skin barrier

↑ Syndecan = good epidermal cohesion = skin resistance and tightness

↑ HA = good hydration and support

Wavy DEJ = attaches the epidermis to the dermis, cells communication

& nutrients exchanges

Aging marker:

Low progerin concentration = no senescence

10

Mature skin

Dermis level:

↓ collagens = loss of tightness

↓ elastin = loss of elasticity

↑ MMPs = fibers degradation

Wrinkles formation Skin sagging & slacking Loss of radiance Skin dryness

Epidermis-DEJ level:

↓ thickness = less resistance

↓ syndecan = less cell cohesion

↓ HA = less hydration & support

DEJ flattening

Induction of syndecan synthesis

Contraction & production of collagen

Inhibition of MMP2, MMP3 & MMP9

Anti-elastase

Reduction of progerin production

Aging marker:

↑ progerin accumulation = senescence

Remodelling effect & wrinkles reduction

Progeline™ Summary Efficacy

Clinical Study #1

Clinical evaluation of Anti-sagging effect

on jawlines

Clinical evaluation of anti-sagging effect on jawlines

Protocol 10 healthy volunteers (females)

Aged between 54 and 64 years old (mean=59);

Having ageing skin with slackening and wrinkles on the jaw lines

Formulation: Emulsion cream containing 2% Progeline™ vs placebo

Application: Each product is applied on split-face and split-neck twice a day (morning & evening)

Times measurement : T0, T+28 days, T+56 days

The study area is the jawlines of the face

Progeline™ decreases significantly the volume of the jaw lines after 28 and 56 days

80% of volunteers had an improvement

Clinical evaluation of anti-sagging effect on jawlines

14 14

Clinical Study #2

Clinical evaluation of skin firmness & elasticity

15

Clinical evaluation of firmness & elasticity

Protocol

13 healthy volunteers (females)

Aged between 54 and 66 years old

Displaying saggy skin on the face (loss of firmness, slackening skin)

Formulation: Emulsion cream containing 2% Progeline™ vs placebo

Application: Each product is applied on split-face and split-neck twice a day (morning & evening)

Times measurement : T0, T+28 days

The measurements are performed on both cheeks (diameter probe = 2mm)

15

16

1. Acquisition of raw data Principle : measurement are taken using the SEM575 cutometer (Courage & Khazaka):

- Creation of a negative pressure on the skin’s surface (300 mbar)

- Measurement of the vertical movement on the induced skin

2. Treatment software and methodology

3 measurements are performed on the study area for each subject and each examination time.

Measurement leads to the determination of 3 main parameters:

- Firnmess = tightness, tonicity

- Elasticity = capacity of the skin to stretch

- viscoelasticity = capacity of the skin to go back to its original state

Measurement are performed on both cheeks : - one side treated with Progeline™ cream - one side with placebo cream

Clinical evaluation of firmness & elasticity Cutometry

16

Progeline™ significantly increases skin firmness, elasticity and viscoelasticity, attesting its efficacy on skin slackening and sagging

88% of volunteers had an improvement

Clinical evaluation of firmness & elasticity

18 18

Progeline™ significantly increases skin firmness, elasticity and viscoelasticity, attesting its efficacy on skin slackening and sagging

D0

Slight improvement

Major building effect over time!

Clinical evaluation of firmness & elasticity

Clinical evaluation of firmness & elasticity

19

Progeline™ clearly reduces wrinkles and skin sagging over time

Clinical evaluation of firmness & elasticity

Progeline™ clearly reduces wrinkles and skin sagging over time

21

Consumer test on overall anti-aging effect

Protocol 13 healthy volunteers (females) aged over 54 with saggy skin on the face

Application: Each product is applied on one randomized split-face and split-neck twice a day

(morning & evening) for 56 days

Progeline™ has 92% overall satisfaction score. Consumers agree that it provides firmer, repulped and denser skin.

Progeline™ Summary Efficacy

Remodelling effect & wrinkles reduction

Tolerance studies

• HET CAM (concentration tested at 20%)

• Acute skin tolerance (single patch test) (concentration tested at 20 %)

• HRIPT (irritation and sensitization) (concentration tested at 20%)

• AMES Test (concentration tested at 20%)

Excellent safety profile

24

Company/

Product

Origin Application In vitro In vivo %

Lucas Meyer Cosmetics Progeline™

Peptide Delay the senescence

process for a firmer skin

↓ progerin ↓ MPPs 2,3 9 and elastase, ↑ collagen lattice contration ↑ syndecan

Tested at 3% (28 days): Jawline improvement: up to -10% +20% of firmness, up to 93% +21% of elasticity , up to 82% +13% of visco-elastity , up to 42% Consumer test: 92% satisfaction

0,5-2%

SEDERMA IDEALIFT™ (2009)

Peptide Gravity control Elastin synthesis (elastin/ Tropoelastin released) +325% on human fibroblasts +14.6 reconstructed skin model +14.8% artificially aged reconstructed skin model

Tested at 4% (2 months) : Firmness +7.8% up to 28% Density +9.2% up to 34% Variation of skin resistance to sagging: up to +24% JAW SURFACE: Image analysis : -11.3% up to -21%

4%

LAB SEROBIO PROTEASYL™

Pisum Sativum (Pea) Extract

Protection and reconstruction of the dermal

network .

Anti-elastase activity in human skin (ex vivo) Booster effect of ECM. Stimulation of collagen type I, III, V and glycosaminoglycans fibroblasts

After only 3 weeks of treatment the cream containing 3% skin elasticity up to 16.7%. Improvement of skin firmness by 35%

3-10%

SEPPIC RETILACTYL D™

(2010)

Black pepper extract

The target solution for firming up supporting

tissues

Adheesion capacity of photoaged fibro by 26%. contraction capacity vs photoaged fibro by 156%. ↑ 20% the migration capacity of photoaged reticular fibroblasts

3% significantly reduces the facial ptosis -8.6% improved skin firmness by 14.3% improved firmness by 8.5% 90% of the volunteers considered that the formula containing had a firming effect.

3%

PROVITAL ZIRHAFIRM™

Zyzipus jujuba seed extract

Restructuring, firming

In vitro test gene expression : most effective actives versus control.

3% to the cheek jaw zone. Skin firmness increase: 14.3% elasticity by 7% Improvement of the facial oval: elevation: 22,5%

2-4%

LAB SEROBIO SYNIORAGE™

Peptide ‘senior’ anti-age targeting the epidermis

Stimulate synthesis of collagen vii (improve adhesion DEJ) Stimulation of syndecan synthesis +130%

Tested at 3% (2 months ): Improvement of skin elasticity & firmness Epidermis cohesion

1-3%

LAB SEROBIO PROTEASYL™

Garden pea

Protection & reconstruction of the dermal

network

Anti-elastase Stimulation of collagen I, III, V, glycosaminiglycans

Tested at 3% (21 days): +14,6% elasticity Tested at 5% (6 weeks): +35% of frimness

3-10%

PRODUCT INFORMATION

INCI NAME Glycerin (and) Water (and) Dextran (and) Trifluoroacetyl Tripeptide-2

ADDITIVE None

APPEARANCE Aqueous limpid solution, odourless and colorless

FORMULATION Should be incorporated at the end of the formulation at a temperature below 40oC

DOSAGE 1-2 %

OPTIMUM PH 4.0-6.0

APPLICATIONS • Anti-aging Care • Skin care for mature skin • Firming Care • Remodelling skin care • Post-pregnancy care

• Men’s care • Body care • Photoaging • Alternative to cosmetic procedures

PRESERVATIVE FREE CHINA COMPLIANT

ECO FRIENDLY

Formulation Tips

• Progeline™ is an clear aqueous solution easy to work and formulate with

• Progeline™ should be added in the cooling phase at the end of the formulating process.

• May provoke Maillard reaction in presence of sugar

• No known incompatibilities.

28

THANK YOU!

2015-12-09

Inhibition of several MMPs

Progeline™ inhibits the main MMPs activities

40

60

80

100

0 0,04 0,125

MM

P-1

act

ivit

y (%

of

con

tro

l)

Progeline™ (%)

Inhibition of MMP-1 activity

-19.4% -8.4%

40

60

80

100

0 0,0005 0,0015 0,005 0,015 MM

P-3

act

ivit

y (%

of

con

tro

l)

Progeline™ (%)

Inhibition of MMP-3 activity

**

-35% -24% -13% -28%

40

60

80

100

0 0,04 0,125 0,375 MM

P-9

act

ivit

y (%

of

con

tro

l)

Progeline™ (%)

Inhibition of MMP-9 activity

** **

-18% -14%

-8%

30

Epidermis action: Stimulation of Syndecan

Progeline™ stimulates syndecan synthesis for a better cell-matrix bonds for a maximal anti-sagging effect

Basal Progeline™ 0,0025%

Protocol • Keratinocytes were incubated for 72h with Progeline™ at different concentrations

• Syndecan-1 was labelled by a specific fluorescent monoclonal antibody and semi quantification was performed using confocal microscopy and IMAGE J software.

Syndecan comes from the root Syndein which mean «To bind together»

Effect of Progeline™ on syndecan-1 synthesis

Flu

ore

scen

ce in

ten

sity

(ua)

0

2000

4000

6000

Basal

Progeline™

0.00025% 0.0025%

+19%

TGF- 10ng/mL

+50% * +56%

*p<0,05

*

Protocol

• Skin explants were pre-incubated in presence or absence of Progeline™ during 30 minutes. Human leucocytes elastase is added for 3 hours.

• Labelling with Orcein to identify elastin fibers in red-brown.

Positive control

Negative control with elastase

Elastin fibers

Elastase + Progeline™ 0.25%

Epidermis

Stratum corneum

Dermis

Inhibition of Elastase Activity on skin explants

Progeline™ reduces the Elastase activity and protects elastin fibers

100% inhibition of

elastase activity

31

Dermis action: Collagen lattices contraction

Protocol • 3D collagen I gels are built in presence of skin fibroblasts and polymerized. Gels are treated with

Progeline™ or positive control (FCS) at day 1 and 2.

• The lattices diameters are measured with a micrometer and the % of contraction was calculated

in comparison with the T0 and the untreated control.

Progeline™ induces collagen lattice contraction to restore a more toned and firm skin

32

0

10

20

30

40

50

60

Positive control (FCS)

0.0025% 0.025%

Latt

ice c

on

tracti

on

(%

) 48%

27% 23%

13% 6%

46%

24h 24h 24h 48h 48h 48h

Progeline™

Effect of Progeline™ on collagen lattice contraction