The 3rd Generation Comthing SGS

The 3rd Generation Comthing SGS Triple Anti-Irritant Complex, Full Recovery

The Formation Mechanism of Sensitive Skin Sensitive skin: remarkably sensitive to external stimuli (Physical and chemical stimuli, allergens, pathogens etc.) When sensitive skin is formed, it is easier to generate inflammation reaction or the skin inflammation already existed. The destruction of skin barrier function

Skin Inflammation Reaction Mechanism Immunocyte

Sophora Flavescens, as the leguminous plants of Sophora genus, is distributed in Russia, Japan, India, North Korea and northern provinces of China, growing within 1500 meters area. Sophora Flavescens root contains a variety of active alkaloids, main ingredients are D-matrine, D-oxymatrine, D-sophoranole, L-Lanagyrine. Sophora Flavescens root extract with strong anti-inflammatory and anti-allergy effects, as a immune inhibitor, reduce the release of allergic medium, is a good reliever for skin itchy symptoms. Ref: Greaf Internal Report. Feb 2016

Glycyrrhiza inflata is known as the king of the medicine, belongs to tonic traditional Chinese medicines. Glycyrrhiza inflata root as a well known ancient chinese medicine, has detoxification, anti-inflammatory and anti-allergy effect, and it is testified by modern pharmacology. Glycyrrhiza inflata contains Glabridin, glycyrrhizic acid, Glycyrrhetinic acid, Glycyrrhetinic alkali, Glycyrrhiza metal salts (calcium iron zinc), liquiritin, asparagine, protein and a variety of amino acids, lipids, mannitol,glycyrrhiza polysaccharide, Glycyrrhiza pigment, fruit acid, microelements such as vitamins etc. Ref: Greaf Internal Report. Feb 2016

Effective ingredients of Glycyrrhiza inflata root contains Glycyrrhizic Acid, which is also called glycyrrhizin, 18 - licorice pavilion Acid. Glycyrrhizic acid is decomposed into glycyrrhetinic acid and Glucuronic acid after absorbed by the body, the former decomposed structure has a strong anti-inflammatory activity, and the second has a strong detoxification effect. Glycyrrhizic acid has two isomers, named 18 - alpha - glycyrrhizic acid and 18 - beta - glycyrrhizic acid.

Scutellaria baicalensis as the labiatae plants of the genus Scutellaria, widely distributed in the southwest,northeast and norththern of China. Its root extract contains many kinds of flavonoids, mainly baicalin, radix scutellariae. Scutellaria baicalensis root extract contains a variety of bioactive flavonoids, which can prevent the release of hyaluronidase, thus has good analgesic action.

The Functions of Comthing SGS Reduces the stimulation, protects the skin barrier Anti-Itching, analgesia, relieving Combines active small molecules with biological polysaccharide, controled- release, long-term action.

Comthing SGS Efficacy Studies HaCat keratinocyte NRU Model SN/T 2328-2009 3T3 Fibroblast cell NRU Model GB/T 21769; OECD129 3D Reconstructed Epidermis Model OECD 439, 431 Closed Patch Test GB17149.2

Efficacy Test: In-vitro Irritation Inhibition Assay HaCat keratinocyte NRU Model Cell level Cell level Seed 96 well Plates Add test sample and SLS, incubation 24h HaCat cell culture Untreated Refresh mediam, add NR, incubation 3h Plater reading OD540nm

Efficacy Test: In-vitro Irritation Inhibition Assay HaCat keratinocyte NRU Model Cell level Cell level 15.6% Reduction SLS (ug/ml) SGS (ug/ml) Result : 50ug/ml SGS can inhibit SLS activity up to 15.6 % in HaCat NRU cell model

Efficacy Test: In-vitro Irritation Inhibition Assay 3T3 Fibroblast cell NRU Model Cell level Cell level Seed 96 well Plates Add test sample and SLS, incubation 24h 3T3 cell culture Untreated Refresh mediam, add NR, incubation 3h Plater reading OD540nm 3T3 cell after NR uptake for 3h

Efficacy Test: In-vitro Irritation Inhibition Assay 3T3 Fibroblast cell NRU Model Cell level 14.5% Reduction SLS (ug/ml) SGS (ug/ml) Result : 12.5ug/ml SGS can inhibit SLS (40ug/ml) activity up to 14.5 % in 3T3 NRU cell model.

3D Reconstructed Epidermis Model

3D RE(Reconstructed) Epidermis Model HS(Human skin) Epidermis in vivo 3D RE HS EpiSkin Bat#15ER270915

3D RE Model Introduction:According to EU cosmetics alternative committee(ecvam),the three-dimensional skin is selected as one of the standard model for test cosmetics raw materials stimulation. Principle:Episkin is an in vitro reconstructed human epidermis from normal human keratinocytes cultured on a collagen matrix at the air-liquid interface. This model is histologically similar to the in vivo human epidermis.the human keratinocytes come from mammary samples obtained from healthy consenting donors during plastic surgery. HIV 1 & 2, B and C hepatitis tests are carried out on the donor bloods as well as verification of the bacteriological & fungal sterility of the cells and absence of mycoplasma. Method:5% SLS as stimulator;test MTT, IL-1α and TNF-α after adding the stimulator and test sample. Test sample:0.2% SGS

Efficacy Test: 3-D Irritation Inhibition Assay 3-D EpiSkin Model by MTT test 3D EpiSkin level Inhibition = 12.7% SLS (10ul) SGS (10ul) Result : 0.2% SGS can inhibit SLS (5%) activity up to 12.7% in 3-D model

Efficacy Test: 3-D Irritation Inhibition Assay 3-D EpiSkin Model by IL-α measurement 3D EpiSkin level Inhibition = 25.3% SLS (10ul) SGS (10ul) Result : 0.2% SGS can inhibit SLS (5%) activity up to 25.3% in 3-D model via IL- α measurement

Efficacy Test: 3-D Irritation Inhibition Assay 3-D EpiSkin Model by TNF-α measurement 3D EpiSkin level Inhibition = 25.6% SLS (10ul) SGS (10ul) Result : 0.2% SGS can inhibit SLS (5%) activity up to 25.6% in 3-D model via TNF-α measurement

Raw264.7 macrophage cell Assay Model Cell level Cell level Materials and reagent: Raw264.7 macrophage cell line, The Chinese academy of sciences Shanghai cell bank of the college of life science Mouse interleukin 6 (IL - 6) ELISA test kit, ml002293 Mouse PGE2 ELISA test kit in mice, ml028719 Method: 1. Day1,Cells were plated at a concentration of 5 x 10 4 cells / well. 2. Day2,Changed the culture medium, LPS 1ug/ml as positive control group, blank culture medium as negative control group. 3. Day4, Take the supernatant,the cell viablilty was tested by neutral red uptake, PGE2 and IL-6 was tested by Elisa test kits.

Efficacy Test: Raw264.7 macrophage cell Assay Inhibition =15.9% LPS (1ug/ml) SGS (ug/ml) 1ug/ml LPS showed significant cytotoxic effect after 48h treatment. SGS can increase 15.9% cell viability at 200 ug/ml,showing its potential anti-inflammatory action.

Efficacy Test: Raw264.7 macrophage cell Assay Cell level Inhibition =26.2% LPS (1ug/ml) SGS (ug/ml) RAW264.7 cell was treated with LPS(1ug/ml)and SGS at the same time. We tested PGE2 amount after 48h, SGS show 26.2% inhibition at 200ug/ml. SGS effectively reduced the secretion of inflammatory cytokines.

Efficacy Test: Clinical Irritation Inhibition Assay Closed Patch Test Cell level Clinical level Wash and dry forearm test skin area, where put on the enclosed patch with irritants Remove patch after 24 h, test skin hemoglobin values with CK. Apply blank and test sample every 24 hours Test erythema hemoglobin values and take photo record after 48 or 72h

Efficacy Test: Clinical Irritation Inhibition Assay Base SGS SGS 2% L=12 Before 48h after Positive Control Base Dex L=5 Before 48h after We tested the results after 48h treatment, 2% SGS showed better Erythema relief effect than ( L=12 ) 0.075% Dex.( L=5)

1.Reliability : A variety of SCI literature documents, Chinese medicine record, specific active matter. 2.Validity : ü 3T3 cell model:14.5% Relief against SLS stimulation at 12.5ug/ml ü HACAT cell model : 15.6% % Relief against SLS stimulation at 50ug/ml ü 3D RE Model:12.7% % Relief against SLS stimulation (MTT); 25.3% inhibition of IL-1, 25.6% inhibition of TNF-a ü macrophage model:26.2% inhibition of PGE2 ü Clinical patch:2% comthing SGS show obvious Erythema relief( Delta L=12) 3.Safety: Chicken embryo HET - CAM experiment: non-irritant Human skin closed patch: non-irritant 4.Convenience : Good solubility in water, recommended in a variety of formula

Comthing SGS The 3rd Generation Controlled-release, Longterm action Glycyrrhiza inflata root and Scutellaria baicalensis root contains a large number of bioactive polysaccharide, these macromolecular polysaccharide can form a elastic biofilm on the surface of the skin, to avoid the direct stimulation of the skin A variety of bioactive small molecules (such as glycyrrhizic acid matrine and radix scutellariae, etc.) over the licorice polysaccharide in the formation of the multilayer structure, when macromolecular polysaccharide components in contact with the skin surface, the activity of small molecules can be released slowly, take long-term anti-allergy effect.

Comthing SGS Full Composition Physical Properties Trade name: Comthing SGS INCI name: Water/Butylene glycol/sophora Flavescens root extract/ Glycyrrhiza inflata root extract/scutellaria baicalensis root extract CAS No.: 7732-18-5/107-88-0/Not Applicable/Not Applicable/ 94279-99-9

Comthing SGS Formulation Application Recommended Dosage:0.5-2.0%(W/W) Guidlines: Emulsion System:added into aqueous phase, stirred evenly, can involved in emulsification process; Gel System:directly added to the system, stirred evenly; Applied products: ü Anti-allergy product ü Body care product ü Whitening product ü After-sun care product

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