TranskinTM is a novel peptide developed by LG Household & Healthcare that enhances skin penetration of protein growth factors. TranskinTM was identified using phage display and conjugated to EGF and hGH. This increased their skin penetration 5-fold and absorption 100-fold while maintaining biological activity. TranskinTM-GFs were non-toxic, non-sensitizing, and non-irritating. Their stability was improved by incorporating them into a microemulsion called TransomeTM. A cosmetic containing TranskinTM-GFs showed improved dermal density, elasticity and wrinkle reduction. TranskinTM-GFs were included in OHUI's The First skincare product.

1. TranskinTM
A novel skin penetration enhancing peptide
for the transdermal delivery of
protein growth factors
Sanghwa Lee
Skin Science Research Center
LG Household & Healthcare

2. 1. Introduction
2. Development of TranskinTM
3. Development of TranskinTM-Conjugated
Growth Factors
4. Safety, stability and efficacy
5. Summary
- Contents -

3. Company Introduction
Founded in 1947, LG Household & Health Care, as the first company of LG Group, is the
Korea’s leading company in Fast Moving Consumer Goods industry.
. Founded in 1947
. Annual sales of 4.3 billion US$ (2013)
. 6,500+ employees (with overseas subsidiaries)
Oral Care
Hair Care
Skin Care
Laundry Products
Paper Products
Cleaning &
Hygiene
WHOO
OHUI
su:m
Soo Rye Han
ISA KNOX
LacVert
Cathy Cat
VONIN
CAREZONE
Beyond
The Face shop
VOV
Health &
Nutrition
Supplement
Vitamins, Health
Drinks & Other
Supplements
Coca-Cola
Fanta
Georgia Coffee
Sprite
Vitamin Water
Nestea
Powerade
Minute Maid
Soonsoo
Etc.
LG-Unicharm
Sanitary Napkins &
Diapers
LG-HAI (USA)
LG-Shanghai (China)
LG Beijing (China)
LG Hangjou (China)
LG-VINA (Vietnam)
LG-Taiwan (Taiwan)
Raw Materials :
Stearic Acid,
Glycerin, etc.
HAITAI Beverage
Bong Bong
SunKist
Sunny10
DK
Blackbean Tea
Home care
Personal Care
Nutri-healthBeverage SubsidiariesCosmetics
LG H&H Structure

4. Company Introduction
Premium
(Prestige)
Mid to High-
end
(Masstige)
Mass
Affordable
VONIN
LacVertSOORYEHANISA KNOX
OHUIWHOO
 LG H&H’s prestige brands are
mainly sold through
department stores and door-
to-door sales
 LG H&H operates retail outlets
that sells its masstige brands.
As of 3Q 2010, there were
1,065 outlets nationwide
 In 2010, LG H&H acquired a
leading mass affordable
cosmetic company (THE FACE
SHOP), strengthening the
company’s offerings in the
mass affordable segment
SU:M
CAREZONE
THEFACESHOP
BELIF
 Specialized skin-
care products
made from high-
quality Oriental
herbal materials,
such as ginseng
 Functional
cosmetics
competing with
foreign brands,
targeted for
younger
generation
 Offers mid to entry cosmetic products
 838 domestic stores and 320 overseas stores as of 3Q 2010
 Launched China operations in 2006. Currently have over 100
stores retail presences in 10 provinces
 Specialized skin-care
products marketed as
“Cosmedical” products
 Offers lab-based skin-
care products
 Targeted 20’s~30’s
 Mid-range version of Whoo,
with traditional flavor
 Targeted mid 20’s~mid 30’s
 Offers specialized
skin-care products
for men
Source: Company filings and website
Core brands Non-core brands
LG H&H offers a Wide Range of Skin-Care and Cosmetics Products
from Mass Affordable to Prestige
Beautiful
 Launched in 2007
with the concept
of fermentation
 Rapidly gaining
market traction
(67% YtoY growth
in 2010 3Q)
 Launched in
August 2010
 Offers natural
herb-based
cosmetic
products
 Focused on
moisturizing
products
 Targeted early 20’s
~ early 30’s
Cosmetics
BEYOND
 Launched in 2005
 Eco-Friendly Brand
with wide range of
products including
skincare, hair, body
and color cosmetics

5. Cosmetic Market leader, offering the best skin science technology
Company Introduction O HUI brand

6. The first regenerating line, applying stem cell technology
in skin care products.
• Collaboration with CHA stem cell institute, which is one of the top stem cell
research centers in the world
• The First contains “Stem Cell Formula” as one of the active ingredients
• Stem cell formula is consisted of selected growth factors and cytokines
secreted by stem cells proved to be helpful to skin regeneration
Company Introduction O HUI The First

7. Introduction
• EGF: Epidermal Growth Factor (M.W. 6k Da)
• hGH: Human Growth Hormone (M.W. 22k Da)
• TGFβ: Transforming Growth Factor (M.W. 25k Da)
• FGF: Fibroblast Growth Factor (M.W. 27k Da)
• PDGF: Platelet Derived Growth Factor (M.W. 30k Da)
• Cell proliferation
• ECM(Extra Cellular Matrix) production
• Wound healing
 Growth factors are used in cosmetic formulations
 Commonly used Growth factors

8. • High molecular weight Protein
• Poor skin penetration, especially across the stratum corneum
• Could not achieve in vivo efficacy as expected by in vitro data
• Use of liposome
▷ Low entrapment efficiency
• Introduction of PTD1)
▷ Works on cell membrane, but not on stratum corneum
Introduction
 Growth factors in Cosmetics - Limitations
 Attempts to enhance skin penetration
 An innovative method needed
1) Protein Transduction Domain

9. Bacteriophage M 13 Library1)
• M.W. of bacteriophage M13 : 1.6 x 107 Da
• Random peptide sequence fused to N-terminal of
gene III coat protein
• Selection of Bacteriophage penetrated real pig skin
• Peptide sequence facilitate the penetration of
bacteriophage could also enhance the
penetration of growth factors
Random peptide
sequence
Development of TranskinTM
 Phage display : Principles and Assumptions
1) http://www.mikeblaber.org/oldwine/bch5425/lect33/lect33.htm

10. • 3 rounds of selection using Franz cell and Porcine skin
• DNA sequence analysis of selected bacteriophage
• Amino acid sequence determination form DNA sequence
• 10 candidate sequences, for example, ‘NGSLNTHLAPIL’
• Named “TranskinTM”
Epidermis
Dermis
Stratum corneum
0.7mm
 Phage display : Selection Procedures
Development of TranskinTM

11.  Characterization of TranskinTM Peptides
G: Neutral B: Hydrophobic R: Hydrophilic Amino acids
Transkin Peptide
NGSLNTHLAPIL
PTD1) Sequences
GRKKRRQRRRPQ
RRRRRRRRR
1) Protein Transduction Domain
Vs.
Development of TranskinTM

12. NGSLNTHLAPIL
ACTGSTQHQCG
TranskinTM (S3-2)
Control peptide
GGGVGVAPGK-FITCPenetrated peptide
+RelativeFluorescence
0
2
4
6
8
10
12
14
16
0 1X 5X 10X
Control S3-2
Enhancer
Peptide
*
*
*
 Skin Penetration of TranskinTM Peptide
Development of TranskinTM

13. Development of TranskinTM-Growth Factors
 Mechanism of TranskinTM
Buffer Transkin
SC
SC
Intact SC layer
 Disrupting SC Layers?
☞ No Disrupting SC layers
• Disruption of Lipids or Proteins in the SC1)
▷ Integrity of SC structure
• Modulation of their organization
▷ Order of SC structure
Lipid
Aqueous
Keratin
Organization of SC lipids
Normal Disorders
• X-ray scattering
• FTIR
• DSC Analysis
Transcelluar Intracelluar
1. FTIR2) Spectrum
2940 2900 2860 2820
Absorbance
wavenumber
182% --- Transkin
--- SPACE
--- Buffer178%
 Modulation of SC organization?
2. DSC3) also shows a different pattern
☞ Increase of the Fluidity of SC
1) Stratum Corneum 2) Fourier Transform Infrared Spectroscopy 3) Differential Scanning Calorimeter Analysis

14. Development of TranskinTM-Growth Factors
 Effect of TranskinTM Treatment on Skin Permeation
0 ㎛ 10 ㎛ 20 ㎛ 30 ㎛
XYZ
Cumulative XZ
FITC4) labelled Peptide (2.2 kDa)
BufferTranskin
SC
ED
DM
SC
ED
DM
TranskinTM could enhance the partitioning of drugs into the SC layer

15. • Expression vector construction of TranskinTM peptide fused to
N-terminal of EGF/hGH
• Expression of fusion protein in E. coli
• Purification of TranskinTM-EGF and TranskinTM-hGH
• Validation of Biological activity and Skin penetration
Scheme of production of TranskinTM-growth factors
Vector construction Host strain setup Purified growth factors
Development of TranskinTM-Growth Factors
 Production of TranskinTM-EGF, TranskinTM-hGH

16. • TranskinTM-EGF, TranskinTM-hGH were validated by N-terminal sequencing
• Same biological activities as their mother forms
EGF TranskinTM-EGF
Development of TranskinTM-Growth Factors
 Biological Equivalence of Transkin-Growth Factors
T-EGF1)
0
Activity(x106IU/mg)
EGF
2.5-
2.0-
1.5-
1.0-
0.5-
1) T-EGF: Transkin conjugated EGF

17. • Expression of Extracellular matrix related genes assessed by Real-time PCR
Development of TranskinTM-Growth Factors
 Biological Equivalence of Transkin-Growth Factors
Hyaluronan synthase 3 Laminin Elastin
EGF
10ng/mL
0
2-
RelativeActivity 4-
Control
6-
EGF
1ng/mL
T-EGF
10ng/mL
T-EGF
1ng/mL
hGH
102ng/mL
0
2-
RelativeActivity
2-
Control
4-
hGH
10ng/mL
T-hGH
102ng/mL
T-hGH
10ng/mL
EGF
10ng/mL
0
2-
RelativeActivity
4-
Control
6-
EGF
1ng/mL
T-EGF
10ng/mL
T-EGF
1ng/mL

18. • 5-fold enhancement of skin penetration
Development of TranskinTM-Growth Factors
 Enhanced Penetration of TranskinTM-Growth Factors
(A)
Amountofprotein(ng/ml)
EGF vs. TranskinTM-EGF
0
50
100
150
200
250
300
350
EGF S3-2 EGF
*
Amountofprotein(ng/ml)
0
100
200
300
400
500
600
hGH S3-2 hGH
*
(B) hGH vs. TranskinTM-hGH

19. • 100-fold increase in skin absorption
Development of TranskinTM-Growth Factors
 Enhanced Penetration of TranskinTM-Growth Factors
200 ppm
treatment
Concentration
in skin
Absorption
rate (%)
Relative absorption
rate (fold)
hGH 0.055 0.028 1
TranskinTM-hGH 6.2 3.1 107
D.ME.DS.C
a b
100㎛
100㎛

20. Development of TranskinTM-Growth Factors
 Safety of TranskinTM-Growth Factors
Sample IC50 Category
TranskinTM-hGH IC50≥ 1% Extremely Low
TranskinTM-EGF IC50≥ 1% Extremely Low
Cytotoxicity (MTT Assay)
Sample Relative Fluorescence Intensity Category
TranskinTM-hGH CD54 RFI ≤ 200, CD86 RFI ≤ 150 Not a sensitizer
TranskinTM-EGF CD54 RFI ≤ 200, CD86 RFI ≤ 150 Not a sensitizer
Allergy Test (h-CLAT Assay)
Sample
Irritation
index
Category
Control (Emulsion Type) 0.05 Not an irritant
TranskinTM-EGF 100 ppm, Transkin-hGH 100 ppm 0.03 Not an irritant
Patch Test

21. Development of TransomeTM
 Idea for Stabilization of TranskinTM-Growth Factors
• Stabilization Tech.
1. Liposome
2. Immobilization
3. Microemulsion
Microemulsion
Transkin-GF1) located
on the surface
w/oo/w
Transkin-GF Lecithin
Water
Surfactant
Hydrophilic
drug
Oil
Hydrogenated Lecithin
1) Transkin-GF: Transkin conjugated Growth Factor

22. Development of TransomeTM
 Preparation of Microemulsion with TranskinTM-GF
Ternary System
S+CoS :
Surfactant+
Co-surfactant
50% water
50% Mix A
C
B
A
O/W
μE2)
S+CoS
Water Oil
60% Oil
40% S-CoS
20% water
80% Mix A
20 nm
mean size
25.2 ㎚
Particle analyze
• Transparent
• Very Stable
• Low viscosity
1) TranskinTM-GF: Transkin conjugated Growth Factor 2) μE: Microemulsion

23. Development of TransomeTM
 Enhanced the Stability and Permeation of TranskinTM-GF
ElastinSyn.Rate(%)
4-
2-
0-
Activity
ElastinSyn.Rate(%)
80-
40-
0-
25°C 45 °C
Stability (8 weeks)
8ⅹ
T-EGF
T-EGF-μE2)
Permeation
Relative
Penetration
150-
100-
0-
127ⅹ
EGF
T-EGF
T-EGF-μE
T-EGF T-EGF-μE
• 8-fold increase the stability of TranskinTM-Growth Factors

24. Efficacy of TranskinTM-GF containing cosmetic preparation
Wrinkle Improvement
Dermal Density
10-
5-
0-
control sample
10-
5-
0-
1.7ⅹ
0W
4W
Improvement(%)
2.4ⅹ
0W
4W
control sample
Improvement(%)
0.25
0.27
0.29
0.31
0.33
0.35
0 w 4 w
Skin Elasticity
*
6
8
10
12
14
16
18
0 w 4 w
*
Ra
R7
* p<0.05
* p<0.05

25. • Peptide sequence ‘N-G-S-L-N-T-H-L-A-P-I-L’ facilitating skin
penetration of protein was developed and named “TranskinTM”
peptide using phage display method
• Transkin peptide was conjugated to EGF and hGH
- 5-fold increase in skin penetration, 100-fold increase in skin
absorption
- maintain same biological activities
- show low cytotoxicity, no allergic potential or skin irritation
• To enhance the stability of TranskinTM- GF, they were incorporated into
microemulsion system, TransomeTM
• Cosmetic preparation containing TranskinTM- GF improved dermal density,
skin elasticity and facial wrinkles
• TranskinTM- GFs were introduced to OHUI “The First” as one of the active
ingredients
Summary

26. Thank You !