Aqua Bio Technology presentation at InnoCos Europe, Paris

1

Aquabeautine XL™

The Natural and Gentle Skin Refinisher

ABT 3

Background 4

Effects 5

Efficacy 9

Safety & Toxicity 19

Sustainability 21

Formulating with Aquabeautine XL™ 22

Summary 25

Appendix: Marketing a Fish Derivative 27

Norwegian company founded by Professor

Bernt Th. Walther (PhD) in 2000, an

embryologist.

Listed on Oslo Stock Exchange (Axess)

since January 2008 (ABT.OAX).

Develops & markets marine raw materials

to the global personal care industry:

Truly novel,

100% natural & sustainably sourced

from the pure and protected

environment of Northern waters,

Proven safety and efficacy,

Patented globally.

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ABT ASA in Brief

4

Salmon hatchery workers in Norway

reported that their hands became soft

and smooth after sorting salmon eggs

despite the prolonged exposure to

cold water.

Research interest: a unique serine

endo-protease, the zonase™ enzyme,

was first discovered in 1997 at the

University of Bergen.

Background

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farmed Atlantic salmon

hatching fluid

The zonase™ helps the larva getting

out of its eggshell by digesting this

tough, fibrous protein structure,

but without harming the delicate

baby fish.

ABT has patented the technology to

extract Aquabeautine XL™ from the

water after the hatching has taken

place and the uses of hatching fluid

in cosmetics.

The baby fish is unharmed in the

process.

Hatching Fluid & Zonase™ Enzyme

Aquabeautine XL™

Zonase™ Effect on Human Skin

Denecker G et al. J Cell Biol 2008;180:451-458

General structure of the epidermis In humans, the zonase™ degrades

corneodesmosomes, specific cell

binding sites in the stratum

corneum, similar to its natural

substrate, and gently removes

dead skin cells without harming

the living skin cells beneath.

The zonase™ is the only known

non-selfdegrading and innocuous

protease.

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Aquabeautine XL™: A Unique Protein Solution

zonase™ protease

structural

proteins

glyco-

proteins

Aquabeautine

XL™

Structural proteins

Proteins and peptides from

the eggshell hydrolyzed by

the zonase™ enzyme

Skin conditioning

Zonase™ protease

Micro-exfoliation

Glycoproteins

Secreted by specific cells

of the fish larva

Anti-ageing effect,

trigger skin repair/

rejuvenation 7

The hatching fluid does not only contain the zonase™ enzyme,

It is a potent mixture of diverse proteins with various skincare properties.

Aquabeautine XL™ has 3 main effects on

human skin:

1. Gentle exfoliation: The zonase™ enzyme

delaminates the outer part of the stratum

corneum.

2. Moisturizing effect: Other active

components penetrate and transport

moisture and nutrition deeper into the

stratum corneum.

3. Rejuvenating effect: The effect is caused

by elevated differentiation of cells into

keratinocytes.

Without Aquabeautine XL™

With Aquabeautine XL™

2 1 3

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Skin models from SkinEthic treated with Aquabeautine XL™ for 6 hours at 30º C, Source: University of Bergen

Aquabeautine XL™ Key Benefits

Source: University of Bergen

Skin models (SkinEthic) stained with dancyl chloride (yellow color) treated with 0%, 1% and 10% of

Aquabeautine XL™ in a cosmetic formulation for 10 hours

1 % Aquabeatine XL™ 10 % Aquabeautine XL™ 0% Aquabeautine XL™

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Exfoliation

Apparent exfoliation effect in a concentration dependent fashion

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Results from immunohistochemistry are yet to come!

Reconstructed Human Epidermis

(SkinEthic), exposed for either 5 minutes,

12 hours, 24 hours or 48 hours to either

200 µl of:

1% Aquabeautine XL™,

5% Glycolic Acid (AHA),

1 mU/ml Bromelain (fruit enzyme),

dH2O (control),

on the stratum corneum side.

After exposure, cultures either fixated in

4% (w/v) paraformaldehyde in PBS at 4ºC

or used in a MTT-assay.

Aquabeautine XL™ vs. an AHA and a fruit enzyme

Comparative Study

Comparative Study: SEM Images after 12h

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Evaluation of effect of chemicals on human skin analogue - Source: University of Bergen

x 400 x 5000 x 15000

1mU/ml Bromelain

Desquamation by digestion of the cells envelope.

Deionised H2O

Control

5% Glycolic Acid

Damaged stratum corneum, skin corrosion.

1% Aquabeautine XL™

Desquamation by shedding of intact corneocytes.

Only the cell binding sites have been degraded.

Gentle micro-exfoliation

Comparative Study: SEM Images after 48h

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x 400 x 5000 x 15000

1mU/ml Bromelain

Fairly smooth skin surface but presence of

rough patches.

Deionised H2O

Control

5% Glycolic Acid

Heavily damaged skin: broken cell envelope

with apparent cell skeleton.


Ultra smooth skin surface, better than control.

Skin smoothing effect

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Comparative Study: Histology after 24h/48h

Minimal alterations in

the stratum basale and

the stratum spinosum.

Destroyed living cells,

including epidermal

stems cells from the

stratum basale. Only

pyknotic nuclei left in a

few places and the

anucleated cells matrix.

Cell proliferation and

differentiation.

Skin rejuvenating effect


24h

5% Glycolic Acid

24h

1mU/ml Bromelain

24h

Control

(untreated)


The stratum corneum is highly hydrated in the 3 cases.

48h

Denser stratum granulosum,

higher concentration of

lamellar granules.

Improvement of skin

barrier function

Redness subsiding in 24 hours and disappeared within 72

hours when treated with a cosmetic base cream containing

Aquabeautine XL™.

Soothing effect

Redness of sunburns on a male test subject - Source: University of Bergen

10 % Aquabeautine XL™


0 h 24 h 45 h 70 h

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Apparent Effect on Sunburn

Preliminary study

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Single center, double blind, placebo controlled clinical trial is being conducted to

evaluate the effectiveness and tolerance of topical skin treatments containing

Aquabeautine XL™ in females with mild to moderate photodamaged facial skin.

Duration :12 weeks, visits scheduled at baseline, Week 2, Week 6 and Week 12.

Min. 90 female subjects, ages 40-65, N≥30 for 3 cells: 0% Aquabeautine XL™

(control), 1% Aquabeautine XL™ , 3% Aquabeautine XL™ in vehicle (skin lotion).

Efficacy assessed on both sides of the face using:

• Visual/tactile grading: clinical scoring for lines, wrinkles, mottled

hyperpigmentation, laxity, clarity and roughness, for objective and

subjective irritation,

• Instrumentation: cutometer and transepidermal water loss (TEWL)

measurements,

• Digital VISIA CR photographs,

• Subject self-assessment questionnaires.

Trial started on January 14th. Final results due in June!

In Vivo Clinical Trial

Skin smoothness and

clarity significantly

improved after 2 weeks.

No irritation (erythema,

burning/stinging,

itching) observed, even

at 3%.

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In Vivo Clinical Trial: Results after 2 weeks

N=102, p<0.0001

Tactile/visual clinical grading on both sides of the face, 0-9 scale

-13%

-17%

-20%

-11%

-13%

-16%

-25%

-20%

-15%

-10%

-5%

0%

Pla

cebo

Aquabeauti

ne X

L™

1%

Aquabeauti

ne X

L™

3%

Pla

cebo

Aquabeauti

ne X

L™

1%

Aquabeauti

ne X

L™

3%

Tactile Roughness Dullness

% v

ari

ati

on c

om

pare

d t

o b

ase

line

Greater improvement of skin

smoothness and clarity, actually

perceived by the panelists.

Questionnaires at 6 weeks reveal that

statistically significant differences

were found for mean scores of

statements about overall appearance,

overall feel, smoother and softer skin

between the placebo and the products

containing Aquabeautine XL™.

Ultra mild active ingredient that

perfects the complexion

Skin becomes smooth and even with a

youthful glow. -16%

-25% -26%

-19%

-29% -28% -30%

-25%

-20%

-15%

-10%

-5%

0%

Pla

cebo

Aquabeauti

ne X

L™

1%

Aquabeauti

ne X

L™

3%

Pla

cebo

Aquabeauti

ne X

L™

1%

Aquabeauti

ne X

L™

3%

Tactile Roughness Dullness

% v

ari

ati

on c

om

pare

d t

o b

ase

line

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N=101, p<0.0001

Tactile/visual clinical grading on both sides of the face,

0-9 scale

Final results after 12 weeks are

yet to come!

Aquabeautine XL™ 3%

Befo

re

Aft

er


Cytotoxicity (157 x recommended concentration)

Human keratinocytes were exposed to Aquabeautine XLTM for 6

and 24 hours using a MTT test, no cytotoxic effect was detected.

Het Cam (in vitro ocular irritation assay) (2600 x

the recommended concentration)

Aquabeautine XLTM at this very concentration was scored as 4.5

out of 12 (Slightly irritant).

Single Patch Test (10 tp / 175 x recommended

concentration)

200 µl of Aquabeautine XLTM was left on the back of adults for 48

hours. Aquabeautine XLTM at this concentration was found very

slightly irritant.

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Safety & Toxicity

HRIPT (Human Repeated Insult Patch Test)

(100 tp/ 50 x recommended concentration)

Using a Marzulli-Maibach method, Aquabeautine XL™ at

this concentration can be considered non-irritating and

non-sensitizing.

Ames Test (mutagenicity) (2600 x

recommended concentration)

A Bacterial Reverse Mutation Test (Ames test) found

Aquabeautine XL™ at this high concentration to be non-

mutagenic and non-promutagenic.

No Antibiotic Activity

15 types bacteria and 2 types fungi were tested.

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Safety & Toxicity

Aquabeautine XL™ is produced as a by-product of

the Norwegian salmon farming industry – no

animals are harmed in the process.

Aquabeautine XL™ is not derived from Genetically

Modified Organisms.

Aquabeautine XL™ is collected at the sole

hatchery by Lygrepollen, at the end of

Eikelandsfjord. The hatchery’s fresh water supply

comes from Håvikvatnet, a secluded lake famed

for the quality of its water and fishing.

If not collected, the proteins in Aquabeautine XL™

would be released into the wild and biodegraded.

All proteins are shown to be biodegradable. Nowhere in

nature are proteins seen to accumulate. Proteins are

constantly recycled as the amino acids are used as

building blocks for new proteins.

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Sustainability

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INCI name: Hydrolyzed Salmon Egg Shell

Extract

Easy to formulate: colorless, odorless.

Non tacky, does not impact the aesthetics

or stability of a formula.

Active in a broader and more skin-friendly

pH range than hydroxy-acids: 5.5 – 9, with

maximum activity at around pH 7.5.

Suitable for a wide range of cosmetics

applications, even for sensitive skins.

Recommended level: 0.5 to 3%.

An exceptionally stable ingredient with a

shelf life in cosmetic products measured to

be several years (enzymatic activity

retained).

Formulating with Aquabeautine XL™

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Supplied as a preservative-free, sterile solution.

Ideally, should be kept frozen to ensure maximum shelf life (>24 months). Storage at 4˚C

is possible for several months. (For short periods of time (e.g. transportation) storage at

room temperature is ok.)

Should be used as soon as the bottle seal is broken. Resealing of the bottle is not

recommended.

Should not be exposed to temperatures exceeding 65°C. Best added at the end of the

manufacturing process, during cooling, at a temperature below 40°C.

Solvent Compatible

Water yes

Glycerin yes

Alcohol Small amounts only. 20% alcohol renders the enzymes inactive.

O/W emulsion Acrylates based

yes

O/W emulsion Xanthan Gum based

yes

W/O emulsion yes


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Aquabeautine XL™ has a prolonged micro-exfoliating effect and is ideal for:

Daily-use leave-on facial products: creams, lotions, toners, etc.,

Overnight treatments,

Cleansers/masks, in combination with other ingredients, for immediate

and delayed results,

Body care products: body lotions, hand creams, lip balms.

Its unique properties makes it particularly suitable for the following segments:

Anti-ageing,

Brightening,

After-sun,

Natural & organic cosmetics,

Cosmeceuticals,

Products for sensitive skin,

Spa products.

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Aquabeautine XL™, a gentle multifunctional ingredient to perfect the

complexion, to get a smooth and even skin with a youthful glow.

Aquabeautine XL™ key features are:

Novel and unique,

Biospecific exfoliant that gently detaches dead cells in the outer

layer of human skin without affecting living cells beneath, proven

to render comparable results to common hydroxy-acids without skin

irritation,

Documented added benefits: moisturization, anti-ageing (skin

rejuvenation) and soothing effect (under certain conditions e.g.

sunburn),

100% natural and biodegradable: designed by nature and derived

from a sustainable source,

Safe and non-toxic: extensive testing by independent laboratories,

Ease of formulation: colourless, odourless and active in a broad pH

range,

Suitable for a wide range of cosmetics applications, even for the

most sensitive skins,

Proven stability for years in products under standard conditions,

unlike common proteases,

Patented globally.

Summary

Aquabeautine XL™ : Simply Better than Competition

Features Abrasives Hydroxy Acids Non-specific

proteases

Aquabeautine

XL™

100% natural

Anti-ageing benefit

Biospecific

Non-cytotoxic

Non-irritating

Stable

Broad pH range Does not impact

aesthetics (color,

odor, texture)

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Marketing a Fish Derivative

Aquabeautine XL™ is derived from

salmon roe or “red caviar” as it is

otherwise known.

Given its high price, caviar is

associated with luxury. In Russia and

other Eastern European countries,

caviar is commonly served during

festive occasions. In Russia, the

concept generally includes both

sturgeon roe (black caviar) and salmon

roe (red caviar), the former not

necessarily superior to the latter.

Salmon roe is called "ikura" in Japan, a

derivative of the Russian "ikra" (caviar)

and is considered a delicacy.

Contact

Sales & Marketing Director: Fanny Coste

Tel: +47 90 40 27 09

E-mail: [email protected]

Documents

Aqua Bio Technology presentation at InnoCos Europe, Paris