Marketing Ephemer

EPHEMER TM
From CELEBRITYTM:
World’s 1st technology of macroalgae cell culture
Sourcing the EPHEMERTM for a long lasting anti-aging effect
BM-0100-slides-EPHEMER-GB-01-April 2015
CELEBRITY™
World’s 1st technology of macroalgae cell culture
EPHEMER™
is the 1st active ingredient for cosmetic application,
worldwide, from macroalgae cell biotechnologies
Innovation :
> Isolation of macroalgae
in cellular form
> Culture
in photo-bioreactors
in cellular form
2
EPHEMER™
Sourcing the EPHEMER™ for a long lasting anti-aging effect
Undaria Pinnatifida
A brown algae,
known as wakame in Japan &
sourced in Brittany by BiotechMarine
Life cycle:
2 morphologic forms
1 > sporophyte
(macroalgae form)
2 > gametophyte:
source of EPHEMER™
(cellular form)
branched filaments
formed of elongated cells © BiotechMarine
3
Sourcing the EPHEMER™
EPHEMER™ is an extract of gametophytes: macroalgae cells

in the lifecycle of the Undaria Pinnatifida brown algae.
Gametophytes:
• are autotrophic cells. They produce organic compounds
from CO2 & minerals, using energy from light.
• only live a few weeks in the ocean, before giving a different form.
• are unstable in the ocean &

not available in large quantities.
The culture of gametophytes,

as a rare & precious type of macroalgae cells,
in a laboratory is a World first for cosmetic applications.
4
Benefits from this technology
Growing gametophytes in the laboratory
Technical interest of Gametophytes
> Gametophytes are stable

in our process in the laboratory
(not in the ocean)
> Gametophytes from Undaria Pinnatifida contain

antioxidants molecules
Marketing interest of Gametophytes
> The Value of an ephemeral stage

in the life cycle of Undaria Pinnatifida.
> Essential role as the Source of Life of

© BiotechMarine & HEOS MARINE
Undaria Pinnatifida, for the survival of the species
5
Extraction method
1 > Extraction solid / liquid

(biomass + solvant)
2 > Extraction liquid / liquid

(triphasic association, allowing migration of lipophilic molecules)
3 > Separation of the phases

> Isolating the lipophilic extract of gametophytes:
- carotenoïds (ex :fucoxanthin),

Fucoxanthin: quality tracer in standard
- chlorophylles (6 different types),
- triterpenes
© BiotechMarine
6
Concept of Ephemeral Art
An ephemeral moment
gives intense &
unforgettable memories
An ephemeral experience,
a brief moment with
a long lasting impact
© BiotechMarine & K. RACCAH

Ephemeral beach art
for the launch of EPHEMERTM,
Britanny, March 2015
7
Concept of Ephemeral Art
Brief Fragile Ephemeral

& & &
Memorable Precious Essential
© Mickael LEVY - FOTOLIA

© FOTOLIA
Achenes crested dandelion Buddhist Mandala
8
Protection of the Genes of EVE
thanks to a protected source of life (gametophytes)
Protection of the energetic source that women

inherit from their mother & transmit to their children
EPHEMER™ protects the genes of Eve

(mitochondrial DNA), inherited from mom*
Mitochondrial DNA is inherited from mom

whereas
Nucleus DNA is inherited from mom & dad
EPHEMER™ protects mitochondrial DNA integrity

+63% Aconitase, a mitochondrial enzyme involved
in cell homeostasis and known for its protective
role towards mitochondrial DNA.
© Fotolia
(after 8 days, versus control in skin explants - histological evaluation)
*The genetic patrimony is located in the nucleus & the mitochondria.

In the nucleus, all genes are equally inherited from mom & dad.
In the mitochondria, genes are inherited from mum.
Mitochondria are the energy factory (ATP) of the cells.
9
Ephemeral & long lasting antioxidant
© BiotechMarine
10
Scientific data
Studies in human skin explants
> Histologic study results (1/3)
• General morphology at D8 (Trichrome’s Masson coloration)

Control Vehicle Vehicle +EPHEMERTM
• Histological targets (immunostaining confirmations)

SIRT-1: to confirm a long-term activation
Catalase: to confirm the antioxidant long-term regulation
Aconitase mt: mitochondrial marker  to ensure no mitochondrial damages
 Qualitative & quantitative evaluations
11
Scientific data
• Immunostainings
> SIRT-1
Vehicle Vehicle + EPHEMERTM
epid.area % effect vs V:
+101%*** p<0.001
12
Scientific data

• Immunostainings
Vehicle Vehicle + EPHEMERTM
Mitochondrial
ACONITASE
epid.area % effect vs V: +63%* 0.01<p<0.05
CATALASE
epid.area % effect vs V: +19%*0.01<p<0.05
13
Clinical results
IN VIVO: antioxidant capacity of the skin after 28 days
> Evaluation of the antioxidant capacity of the skin

FRAP method (Ferric Reducing Antioxidant Parameter)
method commonly used in measuring antioxidant power from fruits
• Study synopsis:
- 20 healthy Caucasian volunteers (men & women), mean age of 37years old
- Randomized application of a 1% EPHEMERTM formulation vs placebo
formulation vs no treatment on 30cm² zones on thighs, twice a day
-1 month treatment
• Parameter:
-Strippings
Evaluation of performed
were the antioxidant capacity
in order of stratum
to collect the skincorneum layers using CorneofixTM
UVA stress
Treatment with EPHEMER™ twice a day
Skin stripping D28 D29 Skin stripping

D0
FRAP evaluation Residual oxidation evaluation
Skin stripping
& lipoperoxidation (lipoperoxidation) after UVA stress
- Study conducted at Farcoderm, Italy, nov./dec. 2014 -

14
Clinical results
IN VIVO: antioxidant capacity of the skin after 28 days
> Evaluation of the antioxidant capacity of the skin
• Results:
Formulation with
1% EPHEMER™
Variation after 28 days
(% versus D0 ) +15%
> After 28 days, EPHEMER™ reinforces the antioxidant capacity of the skin
+15% observed versus D0 (p<0.001)
versus the placebo (p=0.005)
versus non-treated zone (p<0.001)
- Study conducted at Farcoderm, Italy, nov./dec 2014 -
15
Better recovering response to oxidative stress
After end of the treatment
% variation D29-D28 of basal lipoperoxidation
On Day 29, once treatment finished,

After UVA stress, lipoperoxidation
treated skin with EPHEMER™
is lower on treated skin with
for 28 days has a better recovering &
EPHEMER™ than on non treated skin
Response to oxidative stress.
Clinical results
IN VIVO: effect on skin relief after 56 days
> Evaluation of skin relief on the crow’s feet region
• Study synopsis:
- 25 healthy Caucasian women, older than 45 years old,

presenting a wrinkle score of 3 to 6 (ProDerm score) on the crow’s feet area
- Randomized application of a 1% EPHEMERTM formulation vs placebo formulation,

twice a day, on half-face
- 2 month treatment
• Parameters:
- Evaluation of the skin relief:
AEVA fringe projection method

(Ra)  skin microrelief
(Rz)  skin roughness, characterizing the average depth of wrinkles
- Study conducted at ProDERM, Germany, nov./dec. 2014 -
17
Clinical results
IN VIVO: effect on skin relief after 56 days
> Evaluation of skin relief on the crow’s feet region

Results: EPHEMER™ decreases the microrelief versus placebo (p=0.08),
Formulation with 1%
EPHEMER™
Microrelief variation after 56 days
(% versus D0) -7% (p=0.02)
Roughness variation after 56 days
(% versus D0) -5% (p=0.02)
Benefits from
EPHEMERTM 1%
on microrelief & roughness
after 56days treatment
D0 D56
EPHEMER™ decreases the microrelief EPHEMER™ decreases skin roughness (-5%),

(-7%) effect observed versus D0 (p=0.02) effect observed versus D0 (p=0.02)
- Study conducted at ProDERM, Germany, nov.-dec. 2014 -

18
EPHEMERTM
Sourcing the EPHEMER™ for a long lasting anti-aging effect
INCI Name:
Caprylic/Capric Triglyceride -
Undaria Pinnatifida Extract
QUALITY TRACER: fucoxanthin

SOLUBILITY: liquid & liposoluble
FORMULATION: easy
PRESERVATIVE: none
RECOMMENDED USE: 1%
APPROVALS:
CHINA COMPLIANT* * * RSPO*

GREEN PALM PROGRAMME
* CHINA COMPLIANT: INCI name listed on IECIC list of June 2014.

* ECOCERT: raw material approved by ECOCERT GREENLIFE, in conformity with Ecocert natural and organic cosmetic standard.
* COSMOS: raw material approved by ECOCERT GREENLIFE, in conformity with Cosmos standard.
* RSPO GREEN PALM PROGRAMME: supports the production of substainable palm oil through the green palm programme (book and claim)
Annex
© BiotechMarine
• Environmental sustainability aspects
• Scientific datas:
Studies on human skin
Environmental sustainability aspects
CELEBRITY™, sustainable technology of macroalgae cell culture
> Protection of macroalgae
EPHEMER™:
- protects the Source of Life (gametophyte) of Undaria Pinnatifida,
- reduces the environmental footprint (local sourcing & manufacturing) & contributes
to the sustainable sourcing of raw material from renewable raw material origin,
- enables a constant supply. No problems related to lack of raw materials, pollution,

or stock,
- process to grow is totally natural. The production is handled in natural sea water.
> RSPO
- SEPPIC is member of RSPO (Roundtable on Sustainable Palm Oil).
- EPHEMERTM is covered by Book & Claim certification (Ephemer supports the production of
sustainable palm oil through the green palm programme)
- SEPPIC intends to get certification for the BiotechMarine Pontrieux plant according to mass Balance
criteria within current year and to propose Ephemer MB (Mass Balance certified) in 2016.
21
Scientific data
Studies in human skin explants - Bio-EC/Genex
+/- daily treatment (2mg/explant):

> Protocol > vehicule (hydroalcoholic gel 2,5%-topic & ethanol 1%-systemic)
> EPHEMERTM (1% in hydroalcoholic gel 2,5%-topic
& 0.1% EPHEMER™ in 1% ethanol-systemic)
24 h D8
Human abdominal Transcriptomic study & Histologic study

skin explant histological quality control
( 10mm) : General morphology
(Masson’sTrichrom coloration)
RNA extraction &
> from a 50year old
caucasian woman
qualification + immunostaining :
(transcriptomic study) Aconitase +
RNA amplification & Catalase + SIRT-1 +
> from a 47year old
hybridization on human
caucasian woman
(histologic study) full genome chip
n=3/condition Relative quantification of Qualitative & quantitative

gene expression evaluation
variation (vs vehicule)
22
Scientific data
Studies in human skin explants - Bio-EC/Genex
> Full genome transcriptomic study results (1/4)

• Histologic quality control at 24h
Control Vehicule (placebo) EPHEMERTM
• Global profile of transcriptomic regulations
Upregulated genes: 635 (fold change ≥ 1.25)

Downregulated genes: 263 (fold change ≤ 0.8)
(results between replicates)
23
Scientific data
Studies in human skin explants (Bio-EC/Genex)

• Some relevant upregulated genes
Upregulated targets (fold change)
HIF-1α 2.61 HSPA4 1.59 MT1E 1.51

Antioxidant
KLF5 2.80 Heat HSPA13 1.38 CAT 1.69
elements
Shock
SIRT1 1.39 AHSA2 1.92 TXNL1 1.69
Proteins
LAMB
ID1 1.99 HSPD1 1.47 2.92
1
Laminins
LAMA
FLG2 2.33 SERPINB3 2.38 1.82
4
• Some relevant downregulated genes

Downregulated targets (fold change)
ROMO1 0.64
COX5B 0.69
Mitochondrial
respiratory COX6A1 0.71
chain NDUFA11 & 13 + B2 & 7 0.67
elements
ATP5E 0.58
24
Ephemeral antioxidant
Mechanism of action: after 24 hours
↘ Free radicals Ephemeral antioxidant

Mitochondria have an ephemeral
ability to produce energy in anaerobic
conditions, leading to a reduction of free
radicals release.
With EPHEMER™
HIF-1α gene expression is up regulated (2.6x more than
control, in skin explants) leading to a switch from aerobic
to anaerobic metabolism.
© BiotechMarine
SIRT-1 gene expression is up regulated (1.4x more than

control, in skin explants) : retrocontrol on HIF-1α for a
balanced regulation of its effects.
25
Long lasting antioxidant
Mechanism of action: after 8 days
> Long lasting antioxidant
EPHEMER™ :
• protects the cells against free radicals
+19% Catalase
(Detoxyfying enzyme – in histologic evaluation)
thanks to SIRT-1 long term activation

SIRT-1 gene expression is up regulated at 24h (1.4x more than control),
regulation maintained after 8 days (+101% Sirt-1 versus control in epidermis staining).
SIRT-1 is also known as the protein of longevity
• protects mitochondrial DNA integrity
+63% Aconitase
a mitochondrial enzyme involved in cell energy production and
also known for its protective role towards mitochondrial DNA.
(after 8 days, versus control in skin explants - histological evaluation)
26
IN VIVO antioxidant protocol
FRAP analysis: study details
(1) Evaluation of the antioxidant capacity of the skin:

FRAP method, ferric reducing antioxidant power
The FRAP method gives information about the global antioxidant power of samples.
It is used:
• in vitro to measure the antioxidant • in vivo on biological samples like human

profile of food products (fruits, plant as well as plasma & skin samples (for measuring the effect of
extracts, etc...) food supplements or topical products)
FRAP is one method to target oxidation in biological samples, as are ORAC, DPPH, ABTS methods…
• 300cm² zones were defined on the thighs for products’ application

• Strippings were performed in order to collect stratum corneum layers using CorneofixTM
• The 1st stripping was discarded; following 2 other strippings were kept frozen at -20°C for further analysis.
antioxidants from stripped

stratum corneum samples
blue colored
Fe(III)-complex Fe(II)-complex
pH=3.6 T=37°C (absorption measured at 595 nm)
27
FRAP analysis: study details
Lipoperoxidation analysis: study details
(2) Evaluation of the cutaneous lipoperoxidation,

24 hours after a UVA stress, through MDA measurement
Malonyldialdehyde (MDA) is one of the main products resulting from lipid peroxidation process.
Its concentration in a biological matrix is a good index of the lipoperoxidation level of the system.
• 300cm² zones were defined on the thighs for products’ application.

• Strippings were performed in order to collect stratum corneum layers using CorneofixTM.
• The first 10 strippings were discarded; stripping #11 was kept frozen at -20°C for further analysis.
MDA NMPI (*) Stable chromophore

From stripped stratum + Added reactive
(absorption peak at 586 nm)
corneum samples compound
(*) N-methyl-2-phenylindole
29
Contacts
Nota
The analytical specifications warranted are only those mentioned on the certificate of analysis supplied with each delivery of the product.
Except as set forth above, SEPPIC* makes no warranties, whether express, implied or statutory, as to the product which is the subject of this document. Without limiting the generality
of the foregoing, SEPPIC* makes no warranty of merchantability of the product or of the fitness of the product for any particular purpose. Buyer assumes all risk and liability resulting
from the use or sale of the product, whether singly or in combination with other goods. The information set forth herein is furnished free of charge and is based on technical data that
SEPPIC* believes to be reliable. It is intended for use by persons having technical skill and at their own discretion and risk. Since conditions of use are outside SEPPIC*'s control,
SEPPIC* makes no warranties, express or implied, and assumes no liability in connection with any use of this information. Nothing herein is to be taken as a license to operate under
or a recommendation to infringe any patents.
* SEPPIC being:
SEPPIC S.A. SEPPIC Castres BiotechMarine Head Office

22 Terrasse Bellini 27, chemin de la poudrerie – BP 90128 Zone Industrielle 75, quai d'Orsay
92806 Puteaux 81105 CASTRES CEDEX B.P. 72 75007 Paris
FRANCE FRANCE 22260 Pontrieux FRANCE
Tel. : +33 (0)1 42 91 40 00 Tél. +33 (0)5 63 72 69 69 FRANCE
© BiotechMarine / BM-0100-slides-EPHEMER-GB-01-APRIL2015
Fax. +33 (0)5 63 72 69 70 Tel : +33 (0)2 96 95 31 32
info.seppic@airliquide.com contact.biotechmarine@airliquide.com
USA - SEPPIC Inc. SINGAPORE - SEPPIC Asia Singapore JAPAN - SEPPIC Japan Office CHINA - SEPPIC China
30, Two Bridges Road, suite 210 3 HarbourFront Place # 09 – 04 Air Liquide Japan's office Room 2909 Nan Zheng Building
Fairfield, HarbourFront Tower Two Granpark Tower 3-4-1 Shibaura 580 West Nan Jing Road
New Jersey 07004-1530 Singapore 099254 P.O. Box 54638 Minato-ku, Tokyo 108-8509 Shanghai 200041
USA SINGAPORE JAPAN CHINA
Tel. : +1 973 882 5597 Tel : +65 6278 6711 Tel : +81 3 6414 6725 Tel : +86 (21) 64 66 01 49
us.seppic@airliquide.com singapore.seppic@airliquide.com japan.seppic@airliquide.com china.seppic@airliquide.com
POLAND - SEPPIC Poland Office DUBAI - SEPPIC Dubai Air Liquide India Holding Pvt Ltd BRASIL - SEPPIC Brasil
CCIF East Wing 5, Block-A, 3rd Floor O2 Commercial Twin Tower Alameda Santos 745/ cj 72 / 7ºfloor
Ul. Widok 8 Dubai Airport Free Zone B/1304, Asha Nagar Road Cerqueira Cesar
00-023 Varsovie UNITED ARAB EMIRATES Mulund (W) 01419-001
POLAND Tel : +971 (0) 4 205 5443 Mumbai 400080 São Paulo – SP
Tel : +48 22 690 68 73 INDIA BRAZIL
Tel : + 91 22 61046464 Tel : +55 11 3242 3911
poland.seppic@airliquide.com dubai.seppic@airliquide.com india.seppic@airliquide.com brasil.seppic@airliquide.com
GERMANY - SEPPIC Gmbh ITALIA - SEPPIC Italia Srl COLOMBIA - SEPPIC Colombia SAS
Martinstrasse 16-20 Via Quarenghi 27 Calle 71 n°10-40
50667 Köln 20151 Milano Edificio Orbe 71 of 401
GERMANY ITALY Bogota
Tel : +49 (0)221-888824-0 Tel : +39 02 38009110 COLOMBIA
Tel : +571 702 44 48
germany.seppic@airliquide.com italy.seppic@airliquide.com colombia.seppic@airliquide.com

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EPHEMER TM

Sourcing the EPHEMERTM for a long lasting anti-aging effect

EPHEMER™ is an extract of gametophytes: macroalgae cells

• are unstable in the ocean &

The culture of gametophytes,

Technical interest of Gametophytes

> Gametophytes are stable

> Gametophytes from Undaria Pinnatifida contain

Marketing interest of Gametophytes

> The Value of an ephemeral stage

> Essential role as the Source of Life of

Undaria Pinnatifida, for the survival of the species

1 > Extraction solid / liquid

2 > Extraction liquid / liquid

3 > Separation of the phases

- carotenoïds (ex :fucoxanthin),

© BiotechMarine & K. RACCAH

Brief Fragile Ephemeral

© Mickael LEVY - FOTOLIA

Achenes crested dandelion Buddhist Mandala

Protection of the energetic source that women

EPHEMER™ protects the genes of Eve

Mitochondrial DNA is inherited from mom

EPHEMER™ protects mitochondrial DNA integrity

(after 8 days, versus control in skin explants - histological evaluation)

*The genetic patrimony is located in the nucleus & the mitochondria.

> Histologic study results (1/3)

• General morphology at D8 (Trichrome’s Masson coloration)

• Histological targets (immunostaining confirmations)

> Histologic study results (2/3)

Vehicle Vehicle + EPHEMERTM

> Histologic study results (3/3)

epid.area % effect vs V: +63%* 0.01<p<0.05

epid.area % effect vs V: +19%*0.01<p<0.05

> Evaluation of the antioxidant capacity of the skin

Treatment with EPHEMER™ twice a day

Skin stripping D28 D29 Skin stripping

- Study conducted at Farcoderm, Italy, nov./dec. 2014 -

> Evaluation of the antioxidant capacity of the skin

- Study conducted at Farcoderm, Italy, nov./dec 2014 -

% variation D29-D28 of basal lipoperoxidation

On Day 29, once treatment finished,

> Evaluation of skin relief on the crow’s feet region

- 25 healthy Caucasian women, older than 45 years old,

- Randomized application of a 1% EPHEMERTM formulation vs placebo formulation,

AEVA fringe projection method

> Evaluation of skin relief on the crow’s feet region

EPHEMER™ decreases the microrelief EPHEMER™ decreases skin roughness (-5%),

- Study conducted at ProDERM, Germany, nov.-dec. 2014 -

QUALITY TRACER: fucoxanthin

CHINA COMPLIANT* * * RSPO*

* CHINA COMPLIANT: INCI name listed on IECIC list of June 2014.

> Protection of macroalgae

- enables a constant supply. No problems related to lack of raw materials, pollution,

+/- daily treatment (2mg/explant):

Human abdominal Transcriptomic study & Histologic study

n=3/condition Relative quantification of Qualitative & quantitative

> Full genome transcriptomic study results (1/4)

• Global profile of transcriptomic regulations

Upregulated genes: 635 (fold change ≥ 1.25)

(results between replicates)

> Full genome transcriptomic study results (2/4)

HIF-1α 2.61 HSPA4 1.59 MT1E 1.51