Unisooth EG-28 Rapid Control of Skin Irritation for the removal of Dark Circles

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Unisooth EG-28 Rapid Control of Skin Irritation for the removal of Dark Circles The appearance of dark circles is a biologically complex process closely linked to subocular micro-inflammation. By actively ling this localized inflammation, Unisooth EG-28 acts on the 3 major causes of dark circles: 1) Reduction of skin inflammation Decrease of redness, itching and pigmentation; 2) Stimulation of lymphatic circulation Reduction of periorbital edema; 3) Vasorelaxation of under eye capillaries Microcirculation improvement.

Focus on the product Under eye dark circles are triggered by irritation Dark Circles are the result of a complex set of biological parameters, that are closely linked to inflammation: Excessive pigmentation (accumulation of melanin) 1,2 directly increases when skin is irritated. Periorbital edema 2 becomes more visible when inflammation increases. Poor blood circulation 3 causes the accumulation of non-oxygenated blood in capillaries and tissues due to blood leakage which increases the dark-bluish color of dark circles. All of these processes increase upon aging and lead to darkening of the skin color under the eyes. Unisooth EG-28: an optimized blend of Gallate derivatives... Unisooth EG-28 is a balanced combination of activated forms of gallic acid which consists of the following: A glycosylated form of gallic acid coming from oak leaves A glycosylated form of epigallocatechin gallate originating from green tea Propyl Gallate While propyl gallate is a well known lipoxygenase inhibitor and antioxidant in literature, gallic acid and epigallocatechin gallate inhibit histamine release, decrease cytokine production and reduce immune cell infiltration. The combination of these substances results in strong anti-inflammatory action coupled with significant antioxidant properties.... to get rid of dark circles by decreasing under eye skin irritation Unisooth EG-28 acts synergistically on several biological targets to block the initial causes of dark circle formation: It inhibits one of the major inflammation pathways of the body, NF-kB. This results in preventing the release of IL-8 and CXCL-1, two of the key cytokines involved in skin irritation and redness. It activates the pro-lymphatic factor VEGFR-3 gene expression to increase lymph circulation and reduce periorbital edema. It activates Heme Oxygenase 1 gene expression which is an anti-inflammatory and antioxidant factor. It is also well known to generate CO with vasorelaxant and endothelial cell protection effects. It assists in the recovery of normal blood flow under the eye and lessens blood leakage. Clinical tests on 25 subjects have demonstrated a significant decrease of skin redness, water loss and appearance of dark circles as soon as 15 days! Under eye dark circle Irritated darkened skin Nf-kB activated: IL-8 and CXCL-1 released Hyperpigmentation due to melanin production Periorbital edema Increased blood pressure Unisooth EG-28 Normal under eye skin Soothed skin Nf-kB inhibited, IL-8 and CXCL-1 release decreased Melanin production diminished Lymph circulation increase Blood pressure decrease and increased blood flow circulation 1. Dermatological Surgery, 29; 35:1163-1171 2. Journal of Cosmetic Dermatology, 27; 6:211-215 3. Journal of Cosmetic Dermatology, 24 ; 3(2):73-5

Biological activity Controlling key inflammation factors (in vitro tests) Unisooth EG-28 was tested for its ability to inhibit the NF-kB pathway. Transformed human cells (HT29) were transfected with a NF-kB reporter gene plasmid. These were incubated for 24 hours with TNF-α in the presence or absence of Unisooth EG-28 at differing concentrations. A colorimetric reaction determined the NF-kB activity. Unisooth EG-28 was tested for its ability to inhibit interleukin 8 (IL-8) and chemokine CXCL1 release in normal human epidermal keratinocytes (NHEK). The NHEK were incubated with TNF-α and interleukin-1α (both at 5 ng/ml, referred to as CTK mix). Unisooth EG-28 was compared to the reference NF-kB Inhibitor III. Control of NF-kB activity Control of IL-8 and CXCL1 release % of induced 1 8 6 % vs untreated 1 8 6 4 Treatment 4 2 Unisooth EG-28.2% Unisooth EG-28.14% Unisooth EG-28 1.1% 2-2 -4 CTK mix CTK mix + 5 um CTK mix +.8% CTK mix +,4% NF-κB Inhibitor III Unisooth EG-28 Unisooth EG-28 IL-8 CXCL1 Result: Treatment with increasing doses of Unisooth EG-28 significantly inhibits up to -85% of the Nf-kB activity in human cells induced with TNF-α. z<1.96 Wilcoxon test Result: Unisooth EG-28 significantly inhibits the release of IL-8 and CXCL1 inflammatory cytokines. The inhibitory activity at higher doses was superior to NF-kB Inhibitor III and up to -99% of IL-8 release and -135% of CXCL1 release compared to CTK mix. p<.1/p<.1 compared to /CTK mix, Student s t Test Promoting lymph circulation and vasorelaxation (in vitro tests) Unisooth EG-28 was tested for its ability to stimulate the pro-lymphatic factor Vascular Endothelial Growth Factor Receptor 3 (VEGFR 3) and the Heme Oxygenase 1 gene expression. Human dermal microvascular endothelial cells were incubated for 24 hours in a medium containing or not containing Unisooth EG-28. Lymph draining factor VEGFR 3 gene expression Vasorelaxant factor Heme Oxygenase 1 gene expression % of 8 6 4 2 % of 3 25 2 15 1 5 Control Unisooth EG-28.4% Unisooth EG-28.2% Control Unisooth EG-28.4% Unisooth EG-28.2% Result:.2% of Unisooth EG-28 increases VEGFR 3 (+675%) and Heme Oxygenase 1 (+194%) genes synthesis.

Efficacy Clinical evaluation: reduction of skin inflammation... Unisooth EG-28 was tested versus a placebo at 1% and 3% for its ability to reduce skin irritation. Skin irritation was induced by a 24h patch occlusion with a 2% sodium lauryl sulfate (SLS) aqueous solution on the skin. Erythema index (Mexameter ) and Trans-Epidermal Water Loss (Tewameter ) were evaluated after 15 min, 3 min, 6 min and 12 min. Result: Unisooth EG-28 significantly reduces Instant skin redness reduction SLS-induced skin irritation compared to placebo. Redness was significantly decreased after 15 min (-8%) and up to -15% after 12 min, compared to placebo. TEWL (data not shown) was significantly decreased after 3 min (-17%) and up to -23% after 12 min, compared to placebo. % vs T 1 5-5 -1-15 -2 15 min 3 min 6 min 12 min p<.5 compared to placebo, Student s t Test Unisooth EG-28 3.% Unisooth EG-28 1.% Placebo Untreated... Leading to a visible reduction of dark circles A clinical trial was performed to evaluate the efficiency of Unisooth EG-28 on the reduction of dark circles. A gel containing 3% of Unisooth EG-28 was tested against a placebo in a double-blind study on 25 human volunteers. The products were applied twice per day for 3 days on the area around the eyes. Color changes of the dark circles were evaluated (spectrophotometer) after 15 and 3 days. Reduction of Dark Circles - Red Component (a) Reduction of Dark Circles - Blue Component (b) Color 3,5 Difference 3,4 (Δa) 3,3 3,2 3,1 3 2,9 2,8 2,7 2,6 2,5 COlor -2 Difference -17% -21% (Δb) -2,2-2,4-2,6-2,8-3 -3,2 T days T15 days T3 days T days -14% -21% T15 days T3 days Placebo Unisooth EG-28 3% Placebo Unisooth EG-28 3% p<.1 compared to placebo, Student s t Test p<.5 compared to placebo, Student s t Test Day Puffiness and Day 3 dark circles Reduced puffiness and erased dark circles Result: Unisooth EG-28 significantly reduces dark circles compared to placebo in only 15 days with a visible reduction of under eye edema. There is a significant reduction of the Red (-21%) and the Blue (-21%) components after 3 days.

Summary Technical information INCI: Origin: Preservation: Appearance: Solubility: Water, Propyl Gallate, Gallyl Glucoside, Epigallocatechin Gallatyl Glucoside Glycosylated plant extract Preserved with Benzyl Alcohol and Dehydroacetic Acid Clear, yellow liquid Water-soluble Dosage: 1 3% Processing: Can be added at the end of the formulation process under stirring or homogenizing. It can be heated for a short time with the water phase of a formulation. Formulate at ph between 5. and 7. and temperature below 4 C. Claims Claims: Applications: Removes under eye dark circles, anti puffiness, soothes instantly irritated skin, improves under eye microcirculation, protection from environmental stress. Eye contour, eye creams, eye serum, BB (Blemish Balm) creams, soothing treatments, protective care, sensitive skins, after sun / sun care products.

Induchem AG Industriestrasse 8a CH-864 Volketswil T +41 44 98 43 33 F +41 44 98 43 3 sales@induchem.com Induchem USA, Inc. 535 Fifth Avenue, Floor 23 New York, NY 117, USA T 212-756-9918 F 212-756-9942 salesusa@induchem.com Induchem (France) SAS 5, place de la Pyramide Tour Ariane Etage 33 La Défense 9 F-9288 Paris La Défense Cedex T +33 ()1 55 68 12 23 F +33 ()1 55 68 1 ventes@induchem.com www.induchem.com This information and all further technical advice is based on our present knowledge and experience. However, it implies no liability or other legal responsibility on our part, including with regard to existing third party intellectual property rights, especially patent rights. In particular, no warranty, whether express or implied, or guarantee of product properties in the legal sense is intended or implied. We reserve the right to make any changes according to technological progress or further developments. The customer is not released from the obligation to conduct careful inspection and testing of incoming goods. Performance of the product described herein should be verified by testing, which should be carried out only by qualified experts in the sole responsibility of a customer. Reference to trade names used by other companies is neither a recommendation, nor does it imply that similar products could not be used. LEAFLET-EG28-3.12-1