PHYTO-ELASTICIZER BG

LICORICE LEAF EXTRACT

Strong active ingredient derived from

plantation grown licorice

Glycyrrhiza glabra (Licorice)

Licorice Farm ‘Rooprun Pty Ltd. in Australia.

Finley

Glycyrrhiza glabra (Licorice)Licorice Farm ‘Rooprun Pty Ltd. in Australia.

Cultivation Harvest

Farm site area: 750 acres

Botanical Name : Glycyrrhiza glabra Linné (Leguminosae)

Product Name : PHYTO-ELASTICIZER BG

INCI Name : GLYCYRRHIZA GLABRA (LICORICE) LEAF EXTRACT

Marker

Genistein

O

OOH

HO

OH

Glycyrrhiza glabra (Licorice)

Proliferative Effect on Human Fibroblast cells

Human fibroblast cells (NB1RGB)

37℃, 5%CO2, 24h

Sample solution

37℃, 5%CO2, 3days

MTT solution

37℃, 5%CO2, 4.5h

10%SDS in 0.01N HCl

37℃, 5%CO2, overnight

Abs. 570nm

Pro

life

ration

(%)

**: p

Clinical Trial on Wrinkles around Eyes (1)

Subjects

Treatment

Application

6 Volunteers between 27 to 48 years old

Before test & once weekly during testing,

replica of 2cm in diameter from the corner of eye was

made

A cream containing 10% of PHYTO-ELASTICIZER BG

was applied twice daily on the area around eye

:

:

:

Conditions

Measurement All the replicas were photographed and image-processed

with Adobe PhotoshopTM system, and the improvement

of wrinkles was determined by the histological score

digitized as brightness.

:

BeforeHg. 230.5

After 32 daysHg.242.6

BeforeHg. 235.2

After 50 daysHg. 243.1

Replica of Typical Cases

More than 60% of subjects showed significant improvement of wrinkle depth withaverage wrinkle depth score : +5.5

0

5

10

15

Diffe

rence

in the h

isto

gra

m

A.KT.K

Test sample

Placebo

Before After 7 weeks

Effect on small wrinkles

PHYTO-ELASTICIZER BG 10%

This test was performed to show how Phyto-Elastisizer BG can improveturnover of stratum corneum through epidermis metabolism activation.

In-vivo Trial: Stratum Corneum Turnover

Promoting Effect

Procedure

The patch with 5% of dansyl chloride (5-dimethylamino-1naphthalene sulfonylchloride; fluorescent compound) was applied on upper arm of all subjectsfor 16 hours.

After skin was stained with dansyl chloride, the luminescence of stratumcorneum was evaluated by UV lamp (UVL-56 Long Wave UV-366NM).

Lotion with Phyto-Elastisizer BG (5%) and placebo were applied twice a dayon stained skin. Application of lotions was performed until theluminescence induced by dansyl chloride was not observed any more.

Judgement

Efficacy of Phyto-Elastisizer BG was evaluated upon the difference in numberof days before disappearance of luminescence (compared to placebo).

R e s u l t

Lotion with Phyto-Elastisizer BG showed improvement in

turnover of stratum corneum.

23

23.5

24

24.5

25

25.5

Phyto-Elastisizer

BG (5%)

Placebo

Average

length of

turnover

(days)

Focus on Ceramide

New data for

PHYTO-ELASTICIZER BG

Barrier function mechanism of Ceramide

Ceramide: This substance exits in the horny layer and surrounds the corneocyte.

It plays an important role in maintaining barrier function and is comprised of

50 % intercellular lipids.

horny

layer

prickle layer

basal layer

basement membrane

granular layer

melanocyte

Ceramide

water

UV-ray,

Allergen

microorganism,

bacteria,virusloss of moisture

UV-ray,

Allergen

microorganism,

bacteria,virus

inflamation

Keratinocyte

Lack of

Ceramide

Skin with good barrier function Skin with low barrier function

Pathway of Ceramide production

Sphingomyelin synthetase

Serin + Palmitoyl CoA

3-keto-dihydro-sphingosine

Sphinganine

UDP-glucose

Ceramide

SphingomyelinGlucosyl ceramide

Phosphatidylcholine

CeramideUDP

Serin Palmitoyl Transferase (SPT)

Sphingomyelinaseb-glucocerebrosidase

Diacylglycerol

Glucosyl ceramide synthetase

prickle layer

granular layer

(in keratinocyte)

horny layer

(out of cornetocyte)

Promotive effect of SPT mRNA Expression

0

50

100

150

Pro

moti

on r

ate

of

SPT m

RN

A E

xpre

ssio

n (

%)

Control

Phyto-elastisizer BG

0.1% 0.4%

108.8 ±3.9

126.8 ±11.6

add test sample

Incubation for 24 hours

Isolation of total RNA

Evaluation of mRNA levels

by RT-PCR*

RT-PCR*: Reverse Transcription-Polymerase Chain Reaction

Normal Human Keratonocyte Cells

Test Method : Result :

Promotive effect of Ceramide Synthesis (3D Skin Model)

add test sample

Incubation for 7 days

Extraction of Ceramide

TLC development

Spray a copper(Cu)-sulfate reagent

Heating 120 degree C, 10min.

3D model of human skin (EPI-200)

Test Method :

The amount of lipid in spot was measured

by KODAK 1D Image Analysis Software

Am

ount

of

cera

mid

e (mg/w

ell)

Control 5%

7.0 ±0.5

Result :

10%

3.8 ±0.4

7.9 ±0.8

* * p < 0.05%

Phyto-elastisizer BG

0.0

2.0

4.0

6.0

8.0

10.0

Clinical study of Ceramide synthesis

Subjects

Application

Treatment

Measurement

10 male and female volunteers

A lotion containing 1% of PHYTO-ELASTICIZER BG and

a placebo lotion were applied twice daily for 28days

to the inner forearm (24mm x 45mm)

On first and last day during testing,

cyanoacrylate resins were dropped on the test site,

a glass slide was put on and then the corneocytes

were collected.

Measure the amount of ceramide extracted

from collected corneocytoes by TLC method.

The index of ceramide synthesis is the ceramide amount (mg)

per 1mg of corneocyte.

:

:

:

:

Conditions

Clinical study of Ceramide synthesis

A photograph of the stripping process using

cyanoacrylate resin

Result of Clinical study of Ceramide synthesis

0

2

4

6

8

10

1% PHYTO-ELASTICIZER BG

Before After

Placebo sample

Before After

Am

ount

of

Cera

mid

e (mg/m

g)

4.96±0.5 5.64±0.5 5.34±0.5 6.91±0.9

13.7%

29.4%

( n =10 )

Product information

Origin :

Scientific Name : Glycyrrhiza glabra L.

Family : Leguminosae

English Name : Licorice

Part used : Leaf

INCI NAME Composition

GLYCYRRHIZA GLABRA (LICORICE) LEAF EXTRACT

BUTYLENE GLYCOL

WATER

Licorice Leaf Extract 0.50 %

1,3-Butylene Glycol 49.75 %

Water 49.75 %

PHYTO-ELASTICIZER BG :