Natural Anti-Irritant

Natural Anti-Irritant Bio-Relaxer Natural Anti-Irritant Catalogue B103, WOOLIMLIONS VALLEY 2nd B-LINE, SANGDAEWON-DONG, JUNGWON-GU, SEONGNAM, GYENGGI-DO, SOUTH KOREA, 462-120 Tel:+82 2 573 8575 Email: szhen@ibst.kr 1

Merits of Natural Anti-Irritant Natural Anti-Irritant 1. Objective irritation like rash and eczema Bio-tranquilizer reduces objective irritation Bio-tranquilizer is subdivided into (BT-W)water part and (BT-O)oil part a. Multifunctional effects Anti-inflammation Anti-oxidation 2. Helps restore skin for better healthier looking 3. Is composed of Natural material/ingredient 2

Core Technologies Natural Herb Extraction (Specific Activity) - Natural UV Protector - Acne, Dandruff, Tinea Pedis - Extraction of active ingredients from plants and the increased activity by bioconversion Stable Formulation of Active Ingredients - Stabilized active ingredients through electro-spinning, micro-fluidization, 3D conformational structure and etc - Extracellular Matrix composition - Vitamin C & proteases & etc.. were stabilized through BST s safety and stability technology 3

Core Technologies Bioconversion - BCX-A (Anti-acnes) was developed through bio-conversion of Chamaecyparis obtusa s leaf extract. - Whitening agent (arbutin modifier) was developed through symbitic fermentation. Cell culture - ADSC (Adipose Derived Stem Cell) Filtrate, which contains many different kinds of active human growth hormone such as EGF, HGF, basic FGF, VEGF and etc. - We named ADSC (Adipose Derived Stem Cell) Filtrate as SMGF (Stabilized Mega Growth Factor) Human protein cloning EGF was cloned to vector and isolated from Sacchromyces cerevisae 4

Contents What is Natural Anti-Irritant? What does Natural Anti-Irritant do? How well does Natural Anti-Irritant perform? 5

What is Natural Anti-Irritation? Bio-Tranquilier 6

What is Natural Anti-Irritant The use of functional cosmetics has been increased substantially, and at the same time, its side effect (irritation) is also growing problems for functional cosmetics users. Stimulus can be evaluated as two types Subjective irritation Objective irritation 7

What is? 8

Natural Anti-Irritant What is? There are two types of making it possible for the manufacturers to pick the products depending on the manufacturers need. For the application in water phase For the application in oil phase 9

What is? Potato Cordyceps Ashitaba Centella asiatica Magnolia Indicated Substance : Asiaticosides 18-beta glycyrrhetinic acid Indicated Substance : BT-W(cordycepin) 10

Natural Anti-Irritant What is? Common name INCI NAME Scientific Name Potato Solanum tuberosum (potato) extract Solanum tuberosum The potato contains vitamins and minerals, as well as an assortment of phytochemicals, such as carotenoids and natural phenols. The potato is best known for its carbohydrate content (approximately 26 grams in a medium potato). The predominant form of this carbohydrate is starch. It offers protection against colon cancer, improves glucose tolerance and insulin sensitivity, lowers plasma cholesterol and triglyceride concentrations, Anti-cancer, anti-aging, Lower triglyceride 11

Natural Anti-Irritant What is What? is Bio-Relaxer Common name INCI NAME Scientific Name Cordyceps CORDYCEPS MILITARIS MYCELIUM E XTRACT CORDYCEPS MILITARIS Cordyceps is a genus of ascomycete fungi that includes about 400 identified species and many yet to be described. All Cordyceps species are endosparasitoids, parasitic mainly on insects and other arthropods; a few are parasitic on other fungi. The Cordyceps have a long history as medicinal fungi.some polysaccharide components and cordycepin have some anticancer activity in preliminary in vitro and animal studies. It has been used to protect the bone marrow and digestive systems of mice from whole body irradiation Anti-cancer, Hypoglycemic efficacy 12

Natural Anti-Irritant What is? Common name INCI NAME Scientific Name Ashitaba Angelica keiskei leaf/stem extract Angelica keiskei Angelica keiskei, more widely known under the Japanese name of Ashitaba. In folk medicine it is claimed to be diuretic, tonic, to improve digestion, and, when applied topically, to speed wound healing and prevent infection. It also improves the digestion system and strengthens the immune system with its symbiotic micro organism content to strike a natural balance. Anti-oxidant and wound healing 13

Natural Anti-Irritant What is? Common name Centella (Gotu Kola ) INCI NAME Scientific Name Centella asiatica leaf extract, Centella asiatica The plant is one of the best means of regenerative therapy and has for centuries been used in the medicine of China and India. According to the ancient records of oriental herbalists, it has been called one of the miracle elixirs of life. It is used as a stimulating and toning remedy, to improve metabolism and in the treatment of bronchitis, bronchial asthma and tuberculosis Wound healing, stimulate metabolism 14

Natural Anti-Irritant What is? Common name INCI NAME Magnolia Magnolia liliflora flower extract Scientific Name Magnolia liliflora Magnolia is a large genus of about 210 flowering plant species in the subfamily Magnolioideae of the family Magnoliaceae. It is only recently that studies have highlighted the amazing health benefits of two of Magnolia's constituents, honokiol and magnolol. Honokiol has an anti-anxiety effect which is complemented by the anti-depressant effect of magnolol. Honokiol and magnolol are both powerful antioxidants (in vitro, up to 1000 times more potent than alpha tocopherol at inhibiting lipid peroxidation). Anti-oxidant 15

Natural Anti-Irritant What Does Do? Anti- itching Anti-allergy Anti-irritation Antiinflammation Bio tranquilizer 1. Relives the inflammation (caused by irritation) by inhibiting immune response. 2. Suppresses the cytokines caused by inflammation. 3. Cures atopic dermatitis by controlling the immune system skin. 16

Natural Anti-Irritant How well does Bio- Tranquilizer perform? Test Data Lab Data 17

Natural Anti-Irritant Human Patch Test Test Method(Human Patch Test) Test material 1. Volunteers 10 volunteers aged 25~55 Bio Tranquilizer Male 7 Female 3 2. Applied Quantity 0.01ml 3. Method The finn-chamber containing 0.1% of SLS + 0.3% DW and 0.1% of SLS + 0.3% of Bio Tranquilizer were applied to a upper arm of volunteers for 24 hours and then was removed the Bio tranquilizer. After 24 hours from removal of the finn-chamber, skin condition was evaluated after 24hours of removal of the Bio tranquilizer.

Natural Anti-Irritant Human Patch Test Material Control (Distilled water) After application Test 1 (SLS 1% + Distilled water 3%) Test 2 (SLS1% + Bio Tranquilizer 3%) Test number Score ++ + ± - ++ + ± - ++ + ± - 1 - - - - O - - - - - O - 2 - - - - O - - - - - - O 3 - - - - O - - - - - O - 4 - - - - O - - - - - O - 5 - - - - O - - - - O - - 6 - - - - O - - - - - - O 7 - - - - O - - - - - O - 8 - - - - O - - - - O - - 9 - - - - O - - - - - O - 10 - - - - O - - - - - O - A. Sample a. Control : Distilled water b. Test 1 : SLS 1% + Distilled water 3% c. Test 2 : SLS1% + Bio Tranquilizer 3% B. Visual grade a. - : Negative reaction b. ± : Doubtful reaction c. + : Weak positive reaction d. ++ : Strong positive reaction 19

Natural Anti-Irritant Positive Patch Test Sodium Lauryl Sulfate(SLS) is anionic surfactant/skin irritant and inflammatory material when contacted with skin. Confirm the efficacy of BT-W 1. Distilled water 2. SLS 1% + Distilled water 3% 3. SLS 1% + BT-W 3% BT-W decreased irritation caused by SLS. 20

Positive Patch Test 2 Mehtod Irritation was induced by attaching 0.5% SLS using finn-chamber for 24 hours. 0.5% SLS and finn-chamber were removed after 24 hours, and the product with natural anti-irritant was applied twice a day(morning and night). skin reaction was relieved after 7 days Control 7 일차 1. 0.5% SLS 1. Control : Basic Lotion 2. BT-W 2% 3. BT-O 2% 4. BT-W + BT-O 2% 21

Natural Anti-Irritant Human Patch Test Materi al Test 1 (Control : Basic Lotion) Test 2 (BT-W 2%) After application Test 3 (BT-O 2%) Test 4 (BT-W 2% + BT-O 2%) Score Test number +++ ++ + ± - +++ ++ + ± - +++ ++ + ± - +++ ++ + ± - 1 O - - - - - - - O - - - - O - - - - - O 2 O - - - - - - - O - - - - - O - - - O - 3 O - - - - - - - O - - - - O - - - - - O 4 O - - - - - - O - - - - - O - - - - - O 5 O - - - - - - - O - - - - O - - - - - O 6 O - - - - - - - O - - - - - O - - - O - 7 O - - - - - - O - - - - - O - - - - - O 8 O - - - - - - - - O - - O - - - - - - O 9 O - - - - - - - O - - - - O - - - - O - 10 O - - - - - - - O - - - - O - - - - - O A. Sample a. Test 1 : Basic Lotion b. Test 2 : Basic Lotion + BT-W 2% c. Test 3 : Basic Lotion + BT-O 2% d. Test 4 : Basic Lotion + BT-W 2% + BT-O 2% B. Visual grade a. - : Negative reaction b. ± : Doubtful or slight reaction and erythema c. + : Erythema + Induration d. ++ : Erythema + Induration + Vesicle e. +++ : Erythema + Induration + Bullae 22

Sensory Skin Irritation Test Evaluation Evaluation method of International Contact Dermatitis Research Group (ICDRG) - ± + ++ +++ Score 0 0.5 1 2 3 Materi al Test 1 (Control : Basic Lotion) Test 2 (BT-W 2%) After application Test 3 (BT-O 2%) Test 4 (BT-W 2% + BT-O 2%) Dermore action +++ ++ + ± - +++ ++ + ± - +++ ++ + ± - +++ ++ + ± - 10 - - - - - - 2 7 1 - - 1 7 2 * - - 3 - Mean Score 10 0.55 0.45 0.15 Grade Strong irritation No irritation No irritation No irritation 23

IL-6 production (ng/ml) Natural Anti-Irritant Anti-Inflammation IL-6 LPS (Lipopolysaccharide) in HMC-1(Human mast cell line). 12 10 8 6 4 2 0 LPS LPS+BT-W 0.25% LPS+BT-W 0.5% LPS+BT-W 1% LPS+BT-W 1.5% Sample IL-6 production started to decrease dramatically from 1% of BT-W. 24

Nitric Oxide(µ M) Natural Anti-Irritant NO Assay Griess assay method Using RAW 264.7 cell by LPS(Lipopolysaccharide) was used to stimulate RAW 264.7 cell. 100 90 80 70 60 50 40 30 20 10 0 LPS LPS+BT-W 0.25% LPS+BT-W 0.5% LPS+BT-W 1% LPS+BT-W 1.5% Sample NO expression stimulated by LPS decreased remarkably from 1~1.5% of BT-W concentration in RAW 264.7 cell. 25

β-hexosaminidase release(%) Natural Anti-Irritant Beta-Hexosaminidase Assay (PPDA was used to as an allergen in RAW264.7) Inhibition of β-hexosaminidase release was respectively as 100, 84.5, 57.0, 43.6, 32.1% in 0, 0.5, 1, 1.5, 2% of Bio Tranquilizer and it was decreased in a concentration-dependent manner. 100 80 60 40 20 0 0 0.5 1 1.5 2 BT-W Concentration(%) BTW can be used as natural anti-histamine. 26

DPPH inhibition(%) Natural Anti-Irritant Antioxidative Effect DPPH Assay of Bio Tranquilizer 100 90 80 70 60 50 40 30 20 10 0 0 0.1 0.5 1 2 BT-W concentration (%) Bio tranquilizer showed anti-oxidant activity by concentration dependent manner. 27

Cell viability(%) Natural Anti-Irritant Safety-MTT Assay Cell viability 120 100 80 60 40 20 0 0 0.5 1 2.5 5 7.5 10 BT-W Concentration(%) Raw264.7 cell survival rate was not rarely changed even 10% of BT-W was added in the rwa264.7 cell. 28

Natural Anti-Irritant Physical Stability Physical stability of was evaluated for 6 weeks. (4 C, room temperature, 45 C) Result Time(weeks) Appearance color ph 1 Liquid Light Brown 4.5 2 Liquid Light Brown 4.5 4 Liquid Light Brown 4.5 6 Liquid Light Brown 4.5 Before BT-W BT-O Room Temperature Room 4 C 45 C Before 4 C Temperature 45 C No effects in 1. Quality 2. Color 3. ph 29

Natural Anti-Irritant Skin Irritation Bio-relaxer Bio-tranquilizer 30

Amount ( Billion in USD) Natural Anti-Irritant Market Size Bio-relaxer Bio-tranquilizer Market size of Hypoallergenic CM(cosmetics market) and its forecast. 12 Atopic Market Forecast y = 0.2832x + 7.7253 R² = 0.9998 10 8 6 4 2 0 2012 2013 2014 2015 2016 2017 2018 2019 2020 Year Source : Korea Institute of Oriental Medicine Currently it is forecasted that the size of Atopic market is increasing at the rate of approximately USD 0.28 billion/year from 2012 to 2020. 31

Improved BT-W( Lot15E01)

Improved BT-W (Lot15E01) Refinement process improvement Same components with the existing BT-W Improved color Improvement of the smell of raw material (natural substance) Improved abirritation effect

Experimental methods Cause irritation: Attach nonwoven fabric (2cmx2cm) where 0.5g of 20% Lactic acid was dropped to Y zone, and remove it after 2 minutes in order to cause irritation. Abirritation evaluation: Drop BT-W (Lot15E01) and the product of B company respectively, by 0.5g onto the same non-woven fabric (2cmx2cm), and evaluate abirritation by attaching it to the left and right side of the Y zone irritated by lactic acid.

Experimental results (Irritation evaluation) Comparison BT-W (Lot15E01) Irritation rating - ± + ++ +++ 8 2 Product of B company Control group 7 1 1 1 6 4