Issuu on Google+

SymHairTM Force 1631 MICROALGAE NATURAL STRENGTH GREATER HAIR

The information set forth herein does include proprietary and confidential information of Symrise and must be treated in confidence and not be used by the recipient hereof unless otherwise agreed to in writing by Symrise.

AGENDA

PRESENTATION

APPENDIX

HAIR LOSS - CONSUMERS & MARKETS

FOCUS ON SCIENCE

Identifying key consumer needs Hair growth disorders Hair loss causes & reasons

Human hair figures & structure Hair growth cycle Hair loss main reasons & female pattern

FROM CONSUMERS NEEDS to MICROALGAE Symrise green technology Microalgae biotechnology

SYMRISE‘S SOLUTIONS Screening strategy & mechanism of action In vivo objectivation

PAGE 3

HAIR LOSS CONSUMERS & MARKETS

PAGE 4

CONSUMER INSIGHTS HAIR LOSS - ALOPECIA Hair: Socio-cultural phenomenon of great significance in every civilization Important beauty criterion or sacred character

Hair loss or Alopecia:

Loss of self-esteem and self-confidence usually associated with healthy looking Psychosocial impact influencing the psychological well-being

PAGE 5

THE IMPORTANCE OF BEAUTY HEALTHY HAIR, NATURAL LOOK Consumers agree that your hair is a defining characteristic of who you are Two consumers out of three around the world are proud of their hair What consumers want the most is healthy hair that look naturally beautiful Healthy is the most popular quality they want their hair to reflect (47% globally)* Natural comes second (36% globally)*

They want to sustain their hair health and vitality *Source: Symrise LE CMI global database 2012

PAGE 6

BEAUTY HAS ITS SEASONS PREPARING HAIR IS CAREFUL Consumers care about their hair 31% use hair treatments & balms at least once a week globally*

Many consumers know seasons and climate can affect the life of their hair Temperature change can impact hair and scalp Humidity as well The wind can also generate or increase problems, as well as carry pollution

The idea of preparing in advance for climate and seasonal changes is wellaccepted Many consumers believe that good products need time to work

PAGE 7

HAIR STRENGTH AND VITALITY CONSUMERS ARE READY TO PAY Thinning, lack of volume and/or hair loss are key problems consumers want solutions for 36% of consumers are ready to pay the most they can afford for solutions that work against hair loss*

This can be for different reasons Beauty, health and vitality, strength

This can be at all ages From young people who want to look attractive and at their best to older ones who are more concerned with aging *Source: Symrise LE CMI global database 2012

PAGE 8

HAIR GROWTH DISORDERS AT THE HEART OF CONSUMER’S LIFE

Affects males and females at any age Higher prevalence for men Androgenic alopecia :95% of all hair loss Hair loss can be permanent or temporary

?

PAGE 9

HAIR GROWTH DISORDERS HAIR LOSS CAUSES & REASONS Human scalp hair growth: Anagen: active hair growth (2-6 years) Catagen: regression (4-6 weeks) Telogen: resting (2-3 months) Exogen: release of dead hair at the end of telogen

Hair growth cycle of a human scalp hair follicle Telogen (2-3 months) Catagen (4-6 weeks)

Exogen

Anagen (2-6 years)

Causes: Imbalance of the hair growth cycle Genetic disposition Natural aging process and / or disease Reasons: Hormonal (DHT) Structural deficiency of hair roots Inflammation and/or deficient scalp irrigation PAGE 10

FROM CONSUMERS NEEDS TO MICROALGAE

PAGE 11

FROM CONSUMER NEEDS E. E TO MICROALGAE Strong demand for innovative and natural solutions based on clearly identified mechanism of action New approach to address disorders of hair growth

SymHairTM Force 1631 Natural active ingredient from micro algae to prevent hair loss Natural strength and greater hair

PAGE 12

ALGAE GENERAL Algae eukaryotes pluricellular

Macroalgae

prokaryotes

unicellular

Microalgae

Cyanobacteria

PAGE 13

SYMRISE GREEN-TECHNOLOGY MICROALGAE BIOTECHNOLOGY Screening and development of a new natural biological hair loss prevention from microalgae Cutech Srl Expertise in ex-vivo skin and ex-vivo hair follicle screening Contact to experts in microalgae cultivation and production

Why microalgae? Sustainable manufacturing process from renewable sources From 25-40 thousand registered microalgae species only few are used commercially so far Microalgal biotechnology is an already used technology to produce e.g. vitamins, carotenoids, polyunsaturated fatty acids, proteins, cosmetics and health foods

PAGE 14

MICROALGAE COMPOSITION Huge biodiversity: Interesting source for common and rare compounds potentially active on human tissue O

Alkenones e.g. heptatriaconta-8E,15E,22E-trien-2-one CO2H

Proteins

Amino acids Polysaccharides

Polyunsaturated fatty acids

Minerals

e.g. docosahexaenoic acid (DHA)

O OH

HO

Carotenoids O

e.g. astaxanthin

HO2C

CO2H

Other pigments e.g. phycocyanin

O

N H

N H

N

N H

O

Cyclic peptides e.g. Hassallidin A

PAGE 15

MICROALGAE SELECTION In the course of our extensive screening, we identified Isochrysis sp. var. Tahitian (T-ISO) Marine water microalgae For decades successfully exploited in aquaculture for supporting the larvae rearing of delicate marine fish Live food particularly rich in highly unsaturated fatty acids and other micronutrients : T-Iso is rich in vit. PP and vit. B61 Indication of low toxicity Known cultivability in aquaculture2

Tahitian Isochrysis is a strain of Isochrysis collectable at Mataiva (Tahiti). This strain has been isolated by K. Haines in 1977 at Mataiva, Tahiti. Ideal candidate for producing high value active ingredients

1 De Roeck-Holtzhauer Y. et al., 1991, Journal of Applied Phycology, 3: 259-264 2 Reitan K.I. et al., 1997, Aquaculture 155: 207-221

PAGE 16

MICROALGAE CULTIVATION OPTIONS

-Allows to produce large quantities of microalgae at low costs -High risk of contamination by other species or protozoa

OPEN PONDS

-Instable, uncontrolled environmental conditions -Only few algal species are suitable to be cultured using this technology -Example: Spirulina platensis (production of 3000 t/a)1

CLOSED PHOTOBIOREACTORS

Depending on the specific strain, microalgae can be cultivated in either open ponds or closed photobioreactors

-Requires higher investment and operating costs -Low risk of contamination by other species -Allows controlling of environmental conditions and stable production -Possibility to cultivate a large variety of microalgae strains -Example: Chlorella vulgaris / beta-Glucan (production of 1.5 t/a)1

1 C. Griehl and S. Bieler, Nachr. Chem., 2011, 59, 942-947

Image source: http://www.cyanotech.com/company.html

PAGE 17

MICROALGAE BIOMASS PRODUCTION

Microalage supplied by Archimede Ricerche Plant

- F&M: Fotosintetica & Microbiologica, Professor M. Tredici, GWP (Green Wall Panel) - Archimede Ricerche, Dr Silvio Mangini

PAGE 18

MICROALGAE BIOMASS PRODUCTION

Sun

Microalgae inoculum

Artificial sea water

CO2

Vitamins

Mineral nutrients

Microelements

Microalgae biomass

Drying

Extraction PAGE 19

SYMRISE‘S SOLUTIONS SymHairTM Force 1631 A NEW, HIGHLY POTENT

HAIR LOSS PREVENTION ACTIVE

FROM MICROALGAE

PAGE 20

SYMRISE’S SCREENING MODELS HUMAN HAIR FOLLICLES Screening strategy for hair loss prevention active: Hair shaft elongation ex vivo organ culture of human hair follicle ex vivo human scalp culture

Mechansim of action ex vivo organ culture of human hair follicle

Objectivation in vivo

PAGE 21

SCREENING EX VIVO HUMAN HAIR FOLLICLES Hair shaft elongation at day 9 Test protocol Human micro-dissected scalp hair follicles obtained from plastic surgery were transferred into 24 well culture plates After18 h of pre-incubation, hair follicles suitable for testing were selected (good vital stage and a growth of not less than 0.2 mm) 12-18 hair follicles per concentration and control were used Test compounds were applied on day 1 and reapplied every second day* On day 9 (after 8 days of treatment), hair follicles were photographed and the elongation of each hair follicles was measured

* Preparation of supplemented media by addition of DMSO dissolved test compounds into the standard culture medium

Micro-dissected human hair follicles

PAGE 22

SCREENING EX VIVO HUMAN HAIR FOLLICLES Test protocol schematically Selection of suitable hair follicles

Photography and analysis of growth performance

Preincubation

Day 0

Treatment (8 days)

1

2

3

4

5

6

7

8

9

Test compound and medium renewal using the prepared supplemented media (= medium with or without test compounds)

PAGE 23

SCREENING EX VIVO HUMAN HAIR FOLLICLES Positive controls / benchmarks in the hair shaft elongation assay

Effect in the hair shaft elongation assay (hHFs)

Literature

Insulin

Stimulation

Stimulation

Cyclosporine A

Stimulation

Stimulation

Minoxidil *

no effect

Contradictory results

Finasteride **

no effect

Contradictory results

Compound

Carnitine tartrate***

* Blood cirulation enhancer

Stimulation

1 2

Stimulation3

Micro-dissected human hair follicles

1. Philpott et al, 1994, J Invest Dermatol 2. Taylor et al, 1993, J Invest Dermatol

** 5-alpha reductase inhibitor

3. Foitzik et al., 2007, Exp Dermatol

*** patented (Henkel)

PAGE 24

SCREENING EX VIVO HUMAN HAIR FOLLICLES BIO1631 identified as highly potent candidate Average hair growth stimulation (based on responding donors) at 0.004 ppm: +13%

Hair shaft elongation in vitro of BIO1631 after 8 days of treatment * +13% 16

Insulin (20 ppm): +9% Carnitine tartrate (0.2 ppm): +11%

Effective concentration in vitro: 0.004 - 0.4 ppm Responsiveness: 73% Number of tested donors: 15 (11 responders)

Average hair growth stimulation [%] based on responding donors

Cyclosporin A (0.2 ppm): +10%

* 12

+8%

+11%

*

+10% +9%

+7% 8

4

0 0.004

0.04 BIO1631

0.4

0.2 20 Cyclosporin Insulin A

0.2 Carnitine tartrate

concentration [ppm]

*

p < 0.01

PAGE 25

PROOF OF CONCEPT EX VIVO HUMAN SCALP MODEL Hair shaft elongation at day 6 Test protocol Human scalp skin samples obtained from plastic surgery were transferred into culture plates Pictures of the skin samples were taken on day 0, 3 and 6 and each hair shaft was labeled and measured at day 0, 3 and 6 BIO1631 formulated in a tonic or placebo was once daily applied topically on the skin samples (3 samples per treatment) The elongation of each growing hair follicle was obtained by calculating the difference in hair shaft length between day 6 and day 0 Hair follicles that did grow less than 50 ÂľM in 3 days were considered to be dead and were not considered

Human scalp culture

PAGE 26

PROOF OF CONCEPT EX VIVO HUMAN SCALP MODEL Results Control

BIO1631 confirmed as excellent hair growth stimulator on ex vivo human scalp skin

4 ppm

40 ppm BIO1631

Formulation: aqueous alcoholic tonic Number of experiments / donors: 2 Mean stimulation versus placebo from both experiments: +46% at 40 ppm BIO1631 +12% at 4 ppm BIO1631

Data from 1 experiment with representative pictures

PAGE 27

MECHANISTIC INSIGHTS BIO1631

PAGE 28

ANTI-HAIR LOSS PROBLEM, CAUSE, SOLUTION Aging

Spontaneous hair loss

Loss of switch telogen / anagen

Telogen Catagen

Exogen

Solution:

Prolongation of the anagen phase Blocking of transition into catagen

Hair cycle

Anagen

Stress

Stress-induced hair loss

Premature entry into catagen

PAGE 29

MECHANISM OF ACTION HAIR CYCLE STAGING Test protocol Human micro-dissected scalp hair follicles obtained from plastic surgery were transferred into 24 well culture plates After 1 day of pre-incubation, hair follicles suitable for testing were selected 12 hair follicles per concentration and control were used Test compounds were applied on day 1 and 3* After 5 days of organ culture, hair cycle staging was performed by morphological analysis of the bulb region stained with haematoxylin-eosin Extrapolation of the hair cycle stage of the examined hair follicle on the basis of the accepted morphologic criteria after 9 days of culture (MĂźller-RĂśver et al., 2001, J. Invest. Dermatol.)

* Preparation of supplemented media by addition of DMSO dissolved test compounds into the standard culture medium

PAGE 30

MECHANISM OF ACTION HAIR CYCLE STAGING Results +27%

Hair cycle staging by morphologic analysis at day 5 of hair follicle organ culture

70%

55% 45%

-33%

Control

30%

0.04 ppm BIO1631

At 0.04 ppm BIO1631 increases the percentage of hair follicles in the anagen phase versus control by 27% PAGE 31

MECHANISM OF ACTION PROLIFERATION / APOPTOSIS Test protocol Human micro-dissected scalp hair follicles obtained from plastic surgery were transferred into 24 well culture plates After 1 day of pre-incubation, hair follicles suitable for testing were selected 12 hair follicles per concentration and control were used Test compounds were applied on day 1* After 3 days of organ culture, TUNEL / Ki67 / DAPI triple immunostaining was performed Quantitative investigation of the effect of the tested compounds on the induction of apoptosis and/or proliferation of the cell populations localized on the bulb region TUNEL and Ki67 label apoptotic and proliferating cell nuclei, respectively DAPI, staining all the cell nuclei, is used as a control * Preparation of supplemented media by addition of DMSO dissolved test compounds into the standard culture medium

PAGE 32

MECHANISM OF ACTION PROLIFERATION / APOPTOSIS Results:

Ratio of proliferation / apoptosis at day 3 of hair follicle organ culture

p < 0.05 Control

0.04 ppm BIO1631

0.4 ppm BIO1631

BIO1631 significantly decreases the number of apoptotic cells by 59% and 63% versus control at 0.04 and 0.4 ppm, respectively

PAGE 33

MECHANISM OF ACTION PROLIFERATION / APOPTOSIS Results II

Representative pictures Proliferation / apoptosis at day 3 of hair follicle organ culture Control

0.04 ppm BIO1631

0.4 ppm BIO1631

Visable decrease in apoptotic cells (green) and increase in proliferating cells (red) versus control DAPI (used as control): all nuclei in blue PAGE 34

OBJECTIVATION IN VIVO HAIR LOSS PREVENTION

PAGE 35

HAIR LOSS PREVENTION CLINICAL STUDY Protocol: 30 male volunteers (age: 18 to 60 years) suffering from alopecia in stages II-IV of the Norwood-Hamilton scale divided into 2 groups of 15 subjects each 1 group applied the active tonic while the other group applied the placebo Active tonic: 4% SymHairTM Force 1631 (= 160 ppm BIO1631) in water 30% and ethanol 66%, placebo: water 34% and ethanol 66% Application mode: once daily approx. 2 ml on the whole scalp for 3 months Reading: Start (T0) and after 3months Assessment criterion: Instrumental phototrichogram analysis: number of anagen and telogen hairs Comparison of a photograph of a defined area of the scalp directly after hair clipping with a photograph taken after a certain period of time long enough to allow hair growth of the clipped segment (48 hours). Grown hairs = in anagen phase , non-grown hairs = in telogen phase

Subjective analysis: Subjectâ&#x20AC;&#x2122;s evaluation of the efficacy of the treatment by filling in a specific questionnaire Clinical analysis: dermatologist judgement of the tolerability of the treatment and of the degree of dandruff, seborrhoea, erythema, itching and burning

PAGE 36

IN-VIVO CLINICAL STUDY PROTOCOL SCHEMATICALLY

Last application Day 84

First application Day 1

Selection of subjects 2 x 15

baseline Day1

- Phototrichrogram analysis - Subjective evaluation - Clinical analysis

Treatment (3 months)

Month1

Month2

Month3

Daily application of Hair Tonic with 4% SymHairTM Force 1631 group 1 versus placebo group 2

PAGE 37

HAIR LOSS PREVENTION CLINICAL STUDY Anagen hair After 3 months of daily treatment with an aqueous ethanolic tonic containing 4% SymHairTM Force 1631 +10.5% !! increase of anagen hairs versus placebo 4.6% enhancement of the number of anagen hair versus start (D0) Placebo: 5.9% decrease in anagen hair versus start (D0)

6

Variation T3months - T0 [%]

Instrumental results Phototrichogram analysis

4 2 0 -2

-6

-5.9%

-8 Placebo

16

+15.1%

14

-11.7% vs placebo

12 10 8 6

+3.4%

4 2 0 Placebo

T0

T12wks

Active

Telogen hair Variation T3months - T0 [%]

Subject 3

+10.5% vs placebo

-4

11.7% !! decrease of telogen hairs versus placebo 15.1% increase in telogen hair versus start (D0) for the placebo compared to only 3.4% increase for the tonic containing the active

+4.6%

Active

PAGE 38

HAIR LOSS PREVENTION CLINICAL STUDY 90

Subjectâ&#x20AC;&#x2122;s evaluation by questionnaire analysis

80% of the subjects who applied the active stated that they perceived Anti hair loss efficacy Stronger and more vital hair Thicker/more voluminous hair

Percentage of satisfaction [%]

After 3 months treatment

80%

80%

80%

80 70

67%

67%

66%

60

Placebo Active

50 40 30 20 10

Dermatologistâ&#x20AC;&#x2122;s evaluation: Very good tolerance after 3 months of treatment

0 Anti-hair loss Stronger and Thicker/more efficacy more vital hair voluminous hair

PAGE 39

SUMMARY EFFICACY Discovery of BIO1631, a new, natural, highly potent anti-hair loss active 13% stimulation of hair growth versus untreated control at 0.004 ppm after 8 days of treatment. Successful proof of concept on ex vivo scalp skin (12 and 42% hair growth stimulation at 4 and 40 ppm, respectively). Decrease of the number of apoptotic cells in the hair bulb and by that increase of the number of hair follicles in the anagen phase were identified as mode of action Proven in vivo efficacy with +10.5% increase anagen hairs !! and -11.7% telogen hairs vs placebo !!  In 3 months, hair appear stronger, more vital, more dense !!

PAGE 40

SymHairTM FORCE 1631 THE DIFFERENCE & USP Natural hair loss prevention

Natural active ingredient from microalgae to prevent hair loss

Naturally

Natural extract from a sustainable and renewable source

Increases of anagen hair Decreases of telogen hair

Cost effective

Mode of action identified :

Dual action benefits:

Decreases significantly the number of apoptotic cells in the hair bulb

Strengthens and thickens hair Prevents thinning and hair loss

Increases the number of hair follicles in the anagen phase

Clinically shown to increase hair density (*vs. placebo)

Easy to formulate (soluble in water, alcohol and glycols)

+10.5% increase of anagen hair 11.7% decrease of telogen hair

Thicker and more luscious hair Improves volume Stronger, healthier more vital hair

TECHNICAL

MARKETING

Prevents hair loss

Patent pending Preservative free

PAGE 41

SymHair® Force 1631 SUMMARY & CONCLUSION ■

Natural hair loss prevention from microalgae

Sustainable manufacturing process from renewable sources

Blend of BIO1631 in pentylene glycol

INCI : Pentylene Glycol, Isochrysis Galbana Extract

Recommended dosage: 4%

Clear to opal black green liquid

Water soluble

Easy to formulate but has to be protected from light

Light sensitive and colored product due to contained chlorophylls and carotenoids

PN #510316

PAGE 42

HAIR SCIENCE BIOLOGY OF HAIR GROWTH

PAGE 43

HUMAN HAIR FIGURES Some facts about hair The full head of hair consists of 120 000 to 150 000 hairs The hair density per cm2 of the scalp is 250 in average The speed at which hair grows is 1 cm per month which corresponds to 0.3 to 0.5 mm per day 12 cm pear year

The number of hairs that we naturally lose each day is between 50 and 100 Keratin is the main constituent of hair Hair is composed of 50% C, 20% O, 17% N, 6% H and 5% S PAGE 44

HUMAN HAIR STRUCTURE

Hair has two separate structures - the follicle in the skin and the shaft that we see Together with the sebaceous gland and the arrector pili muscle, the hair follicle is part of the pilosebaceous unit The hair folicle is a complex mini-organ of its own

Hair growth results from the proliferative activity of matrix keratinocytes which give rise to the hair shaft and the inner root sheath Nutrition of the papilla and the overlying matrix cells is provided via the capillary loop located within the dermal papilla

PAGE 45

HUMAN HAIR GROWTH CYCLE Human hair growth Scalp hair grows in cycles

Hair growth cycle of a human scalp hair follicle Telogen

Each cycle consists of 3 phases Anagen: active hair growth (2-6 years)

Catagen

Exogen

Catagen: regression (4-6 weeks) Telogen: resting (2-3 months) Exogen: release of dead hair at the end of telogen

Typically, 90-85% of scalp hair are in anagen and approx. 10-15% in telogen Normally each hair follicle cycles independently, Thus, the total number of scalp hairs remains stable

Anagen PAGE 46

HUMAN HAIR GROWTH CYCLE

Hair Growth and Disorders

PAGE 47

HUMAN HAIR GROWTH CYCLE Each hair follicle undergoes 10 to 30 hair growth cycles per lifetime The cyclic transformations are controlled by finely tuned changes in the local signaling milieu, based on change in the expression of cytokines hormones neurotransmitters their receptors transcriptional factors enzymes

which act via endocrine, paracrine and autocrine routes Imbalances lead to hair growth disorders PAGE 48

A DYSFUNCTION: HAIR LOSS 3 MAIN REASONS

Androgenic alopecia Hormonal: Structural deficiency of hair roots: weak implementation  effect of loss of extracellular matrix (ECM) Inflammation & deficient scalp irrigation

5 Îą reductase Testosterone Dihydrotestosterone (DHT)

Miniaturization of follicles & regression of hair roots

Hair thinning / shorter, finer hairs / baldness

PAGE 49

A DYSFUNCTION : HAIR LOSS FEMALE PATTERN Female pattern hair loss (FPHL) is also an androgenic alopecia it is the most common type of alopecia : 20% of women in their 50’s have alopecia. The early onset is similar to male pattern hair loss while the late onset appears with menopause. Causes : Androgens : Dehydroepiandrosterone sulfate (DHEA-S) is converted into DHT This mechanism requires 3 enzymes (17β-hydrosteroid dehydrogenase, 5α-reductase and 3β-hydroxysteroid deshydrogenase. Most of the FPHL are androgen-dependent, patients treated with anti-androgens or 5α-reductase inhibitors show increased hair growth.

Hair Growth and Disorders

PAGE 50

A DYSFUNCTION : HAIR LOSS FEMALE PATTERN Estrogens : Hair follicles have estrogen receptors (ERα and ERβ) The precursor Andrestenedione requires 3 enzymes : the 17-hydroxysteroid deshydrogenase, Aromase and the 5αreductase, to be converted in DHT. How estrogens influence FPHL has not been determined yet. Genetics : Endocrine reviews.

This is a polygenic disorder but the genes involved have not been found. Others : Blood circulation Stress, chemotherapy, malnutritionV PAGE 51

SymHairTM FORCE 1631 NATURAL STRENGTH for GREATER HAIR SUMMARY Thinning hair and hair growth disorders are a wide spread global issue that can affect both sexes at any age Anti hair loss actives are proven agents for preventing, slowing or treating hair loss. Discovery of BIO1631, a new, natural, highly potent anti-hair loss active 13% stimulation of hair growth versus untreated control at 0.004 ppm after 8 days of treatment. Successful proof of concept on ex vivo scalp skin (12 and 42% hair growth stimulation at 4 and 40 ppm, respectively). Decrease of the number of apoptotic cells in the hair bulb and by that increase of the number of hair follicles in the anagen phase were identified as mode of action Proven in vivo efficacy with +10.5% increase anagen hairs !! and -11.7% telogen hairs vs placebo !!  In 3 months, hair appear stronger, more vital, more dense !! PAGE 52

SYMRISE, ALWAYS INSPIRING MOREE

DISCLAIMER Symrise makes no warranties, either express or implied, as to the accuracy or completeness of the information set forth herein. Symrise expressly disclaims any implied warranty of merchantability and fitness for a particular purpose. Prospective users are requested to determine for themselves the suitability of Symrise materials and suggestions for any use prior to their utilization. Any necessary approvals from regulatory authorities for finished products must be obtained by the prospective user. Suggestions for applications involving our products or the reference to, or incorporation of, descriptive materials from patents and the citation of specific patents in this document may not be understood as recommendation for the use of Symrise products in violation of any patent or as a permission or licence to use any patent of Symrise or a third party.

PAGE 53


SymHair™ Force 1631 PDF