REGU®-SCENCE Delays the appearance of skin aging by decades*

* in vitro test performed on fibroblast monolayers

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Agenda

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1 Skin ageing prevention – A growing trend

3 REGU®-SCENCE – Unique activity profile

4 REGU®-SCENCE – Your inner beautician

2 Autophagy – A key process to delay senescence

5 REGU®-SCENCE in a nutshell

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Skin ageing prevention - A growing trend in Anti-Ageing Beauty Care

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*Source: GNPD, 2008-2012, “skin ageing prevention”, category skincare

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Skin ageing prevention Selected market examples

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“conceals, hydrates and helps prevent premature signs of aging”

“helps prevent the appearance of future ageing while diminishing the early signs”

“prevents and repairs signs of ageing and promotes skin renewal”

“increase in collagen production and prevention of future signs of ageing”

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Start early with prevention to delay visible aging!

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• Your skin starts aging before you see it (e.g. collagen breakdown)

• The first signs of aging become visible only over the years

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Agenda

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1 Skin ageing prevention – A growing trend

3 REGU®-SCENCE – Unique activity profile

4 REGU®-SCENCE – Your inner beautician

2 Autophagy – A key process to delay senescence

5 REGU®-SCENCE in a nutshell

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No way not to age!

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• Aging is a natural process - on cellular level it leads to a condition called senescence

• Aging strongly defines how you look • Senescence is accelerated by environmental influences • Senescence is independent from skin types and ethnicity

Both sisters are ageing, but in different ways:

Ageing is due to genetic and environmental factors.

Source: Gunn DA et al (2009), Why some women look young for their age, PLoS ONE 4(12): e8021

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Autophagy process A detailed overview

Autophagy is a very efficient process that not only cleans cells of waste but also regenerates cellular functionality.

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Autophagy process A detailed overview

An essential biochemical housekeeping process

Autophagy is the key process by which cellular waste is removed or recycled. The process starts with the formation of double membrane structures (phagophores). The phagophores then close and form autophagosomes, which sequester damaged cell components. Each autophagosome fuses with a lysosome to form an autolysosome, in which waste matter is degraded by lysosomal enzymes into amino acids, fatty acids, sugars, and nucleotides. These newly formed molecules are now liberated from the autolysosome and can be used for the neo-synthesis of functional biomolecules and for the generation of ATP (improved energy supply).

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Agenda

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1 Skin ageing prevention – A growing trend

3 REGU®-SCENCE – Unique activity profile

4 REGU®-SCENCE – Your inner beautician

2 Autophagy – A key process to delay senescence

5 REGU®-SCENCE in a nutshell

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In vitro screening on key autophagy marker

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LC3 Protein: a widely used marker for autophagy

• Involved in autophagosomes formation & function

• Present on the cells during the whole autophagy process

Screening parameters

• in vitro screening for autophagy induction* with 21 pre-selected substances

• Human HeLa cell line expressing autophagy

• Marker LC3 coupled on gene level to GFP (Green Fluorescent Protein)

• Determination of the fluorescence intensity

Autophagosome visualization by GFP-LC3 fluorescence **

* in vitro screening performed by Centro de Investigaciones Biologicas Madrid (CSIC) ** image by courtesy of P. Boya CIB-CSIC, Madrid

Autophagosomes

Nuclei

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Screening best candidate

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in vitro determination* of the fluorescence intensity increase after incubation with Navarra Asparagus extract:

Untreated Treated with 100 ppm Navarra Asparagus Extract

0.4% of Navarra Asparagus Extract significantly stimulates LC3, a key marker of autophagy, by 29%

* in vitro screening performed by Centro de Investigaciones Biologicas Madrid (CSIC)

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Navarra Asparagus Extract age-dependently stimulates in vitro autophagic activity

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• Method*: in vitro immunofluorescent detection of autophagic activity (LC3 protein) in human dermal fibroblasts

• Treatment: incubation with 100 ppm dry Navarra Asparagus extract for 24h

Decrease of autophagic activity in cells from aged donors

Cells from aged and young donors show similar autophagic activity

* in vitro screening performed by Centro de Investigaciones Biologicas Madrid (CSIC)

Navarra Asparagus Extract

28 years 62 years 28 years 62 years

untreated

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Navarra Asparagus Extract almost completely protects protein oxidation [1/2]

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• ex vivo assay with human skin explants (50 years old donor)

• Cells treated with 3% Navarra Asparagus Extract

vs. untreated cells

• UV-irradiation after 5 days: UVA: 18 J/cm2

UVB: 0.6 J/cm2

• Analysis 1 & 4 days after irradiation

Autophagy decrease in aging skin leads to cellular stress increase, responsible for protein oxidation

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Navarra Asparagus Extract almost completely protects protein oxidation [2/2]

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UV irradiated skin Treated with 3% REGU®-SCENCE 4 days after irradiation

Non-irradiated skin Untreated

epidermis

dermis

• Immuno-staining (dark purple coloration) of oxidized dermal proteins on frozen skin sections

• Irradiation: 18 J/cm2 UVA and 0.6 J/cm2 UVB

UV irradiated skin Untreated 4 days after irradiation

Navarra Asparagus Extract protects irradiated skin from protein oxidation

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As a consequence, Navarra Asparagus Extract reduces in vitro senescence in aged-skin cells

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• Method*: Senescence is monitored in vitro by beta-Galactosidase staining (bluish color, see arrows) in human dermal fibroblasts

• Treatment: incubation with 100 ppm dry Navarra Asparagus extract for 24 h

Increased staining for senescence in cells from aged donor

Decreased number of cells from aged donor becoming senescent

* in vitro screening performed by Centro de Investigaciones Biologicas Madrid (CSIC)

Navarra Asparagus Extract

28 years 62 years 28 years 62 years

untreated

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REGU®-SCENCE – in vivo efficacy Study design

Volunteers: 31 female volunteers, 40 – 55 years old

Formulation: Placebo controlled study

3% REGU®-SCENCE in a cosmetic emulsion

Application: Hemi-face twice per day

56 days

Parameters: - SIAscope measurement (to quantify collagen density)

- 3d ultrasound scanning (to quantify dermis thickness)

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REGU®-SCENCE significantly increases collagen density in vivo

After 56 days, 3% REGU®-SCENCE

• Significantly increases collagen density by 7.5%

• Best individual increase: 61% compared to Day 0

• 4-fold improvement compared to placebo

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REGU®-SCENCE significantly increases dermis thickness in vivo

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After 56 days, 3% REGU®-SCENCE

• Significantly increases dermis thickness by 18% • Best individual increase: 69% compared to Day 0 • Significant improvement compared to placebo

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An unique activity profile Summary [1/2]

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in vitro

• 0.4% Navarra Asparagus Extract significantly stimulates autophagy key marker LC3 by 29%

• The rate of autophagy in fibroblasts from aged donors is increased until levels are similar to those seen in cells from young donors

• It drastically decreases the number of cells from aged donor becoming senescent

ex vivo

• 3% Navarra Asparagus Extract almost completely protects irradiated skin from protein oxidation 4 days after irradiation; significant results already after 1 day

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An Unique Activity Profile Summary [2/3]

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Additional results ex vivo • 4 days after UV irradiation, 3% Navarra Asparagus Extract

significantly increases total collagen network by 5%

• Additionally significantly increases epidermal thickness by 16%

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REGU®-SCENCE Unique Activity Profile Summary [3/3]

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in vivo - 3% REGU®-SCENCE

• Significantly increases collagen density by 7.5% after 56 days; 4-fold improvement compared to placebo

• Significantly increases dermis thickness by 18% after 56 days

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Agenda

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1 Skin ageing prevention – A growing trend

3 REGU®-SCENCE – Unique activity profile

4 REGU®-SCENCE – Your inner beautician

2 Autophagy – A key process to delay senescence

5 REGU®-SCENCE in a nutshell

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• Carefully purified standardized plant bioactive made exclusively from Navarra Asparagus officinalis ‘Grolim’.

• Independently of age, stimulates skin’s autophagy capabilities - the skin key detoxifying process - delaying cellular aging by decades.

• can provide an effect of overall rejuvenation of skin’s appearance.

From 62 back to 26*

* in vitro test performed on fibroblast monolayers

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Spanish Navarra Region The source of exceptional high-quality Asparagus

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The region • Located in northern Spain, bordering the Basque country

and La Rioja • Famous throughout Spain for the high quality of its wine

and food products, incl. ‘piquillo’ peppers and asparagus • Centuries-long Asparagus culture tradition

Optimal culture conditions • Incredibly fertile soil • Ideal spring climate, with cold nights below 12°C and warmer days,

essential for high qualitative products

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Spanish Navarra Region The source of exceptional high-quality Asparagus

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Strict procedures

• Manual harvest before sunrise enables preservation of product quality

• Delivery the same day to the factory for drying, in order to avoid any degradation of the active ingredients

• More than 75 manipulation and manufacturing audits, and around 340 physicochemical analyses routinely performed each season, in order to guarantee highest quality

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Navarra Asparagus The most famous one

26 * source: A. Schwarzbach, Effect of cultivars and deep freeze storage on saponin content of white asparagus spears, Eur Food Res Technol (2006), 222:32-35

• Navarra Asparagus officinalis ‘Grolim’ (D.O. Navarra Spain) contains particularly high quantities of saponins*, known to be positive modulators of autophagy

• Navarra Asparagus contains the saponins Dioscin and Yamoscin as major components

• Saponins are composed of a sugar part and a sapogenin part; the saponins content is measured by means of HPLC after hydrolysis, using a sapogenin standard

• Major sapogenins in saponins from Navarra Asparagus are Diosgenin and Yamogenin

Diosgenin Yamogenin

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Agenda

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1 Skin ageing prevention – A growing trend

3 REGU®-SCENCE – Unique activity profile

4 REGU®-SCENCE – Your inner beautician

2 Autophagy – A key process to delay senescence

5 REGU®-SCENCE in a nutshell

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Delays the appearance of skin’s aging by decades*

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Unique Product Features • Stimulates skin’s own autophagy capabilities, slowing the process

of cellular senescence*

• In vivo significantly improves collagen density and dermis thickness

• Ex vivo reduces UV-induced protein oxidation*

• Made specifically from Navarra Asparagus officinalis ‘Grolim’, which has a particularly high active molecule content

Benefits • Boosts skin’s innate strength to delay the appearance of skin’s

aging by decades • Stimulates skin’s detoxifying processes, independently of age • Provides an overall skin rejuvenation effect REGU®-SCENCE

Raw material approved by ECOCERT GREENLIFE, conform to the ECOCERT Natural and Organic Cosmetics Standard

* in vitro test performed on fibroblast monolayers

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REGU®-SCENCE Your inner beautician

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Cosmetic application • Skin aging prevention for 25+ • Rejuvenating intense care for 40+ • Multifunctional beauty care products, such as BB or CC

Guide formulations with 3% REGU®-SCENCE • “Youth preserving care”: Light, slightly perfumed cream-gel

formulated for young people 25+ who want to take early control of the aging process

• “Cell rejuvenating intense care”: Rich O/W cream with SPF 20 formulated for consumers 40+ who want to delay visible aging by triggering skin’s innate strength

• Samples available on request

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REGU®-SCENCE Regulatory Status

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• Global Status (excl. China): REGU®-SCENCE may be globally commercialized. See specific statement for more details.

• China Status:

Chemical notification Chinese INCI name

2004 CTFA inventory

SFDA list pending for the asparagus extract

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REGU®-SCENCE Formulation guidelines

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Formulation

• Can easily be processed warm (<40°C) during cooling phase • Recommended concentration: 3% • pH stability: at least 3-8 (test range) • Formulation guidelines available under request

Product form • Yellow to light brown hydrophilic solution • INCI name (active): Asparagus officinalis Stem Extract • Standardized in saponins • Formulated with glycerin and propanediol from vegetal origin • Preserved with Potassium Sorbate and Sodium Benzoate /

Calcium Gluconate / Gluconolactone

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Application Concepts

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• Skin ageing prevention for 25+ (with ELHIBIN® PF)

• Rejuvenating intense care for 40+ (with SYN®-COLL)

• Natural rejuvenating intense care for 40+ (with COLHIBIN PF)

• Night cellular rest & detoxifying balm (with DISMUTIN® PF)

• Ultimate skin protection (with ALPAFLOR® EDELWEISS)

• Essential CC Cellular Care (with PEPHA®-PROTECT)

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Disclaimer

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Copyright © 2013 DSM Nutritional Products, 4002 Basel, Switzerland. www.dsm.com/personal-care. All rights reserved. Although DSM has used diligent care to ensure that the information provided herein is accurate and up to date, DSM makes no representation or warranty of the accuracy, reliability, or completeness of the information. This presentation only contains scientific and technical information for business to business use. Country or region-specific information should also be considered when labeling or advertising to final consumers. This publication does not constitute or provide scientific or medical advice, diagnosis, or treatment and is distributed without warranty of any kind, either expressly or implied. In no event shall DSM be liable for any damages arising from the reader’s reliance upon, or use of, these materials. The reader shall be solely responsible for any interpretation or use of the material contained herein. The content of this document is subject to change without further notice. Please contact your local DSM representative for more details. All trademarks listed in this brochure are either registered trademarks or trademarks of DSM in The Netherlands and/or other countries unless explicitly otherwise stated.

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Back up REGU®-SCENCE

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REGU®-SCENCE Particular origin counts for activity

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REGU®-SCENCE outperforms Chinese Asparagus officinalis on Collagen-I synthesis

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SIAscope Principal

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Spectophotometric intracutaneous Analysis Non-invasive method

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Guide formulation “Youth Preserving Care”

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Guide formulation “Cell Rejuvenating Intense Care”

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Guide formulation in vivo study formulation

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