The Science of Staying Healthy

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1 The Science of Staying Healthy Signum Dermalogix, Inc. The Science of Staying Healthy

2 ignum Dermalogix is a spin out of Signum Biosciences, a private biotech company founded in 2004 based on technologies developed by Dr. Jeffry Stock at Princeton University. In 8+ years, we have developed in vitro assay systems and robust analytical expertise including safety, efficacy and claim substantiation for cosmetics, nutraceuticals and pharmaceuticals. We specialize in producing customized cosmetic claim support and/or efficacy dermatological tests providing robust scientific data to support advertising or packaging claims. In addition, we can provide thorough safety testing for your molecule and determine its dermal penetration profile, stability and pre formulation compatibility via our analytical support team (HPLC, LC MS and NMR). Our expertise is also based on standard validated assays in order to assess your molecule/ingredient in well recognized tests. Signum Dermalogix is now bringing these assays and services to you the consumer through this catalog which describes in more detail: Standard assay systems Compound profile screening Analytical services Customized screening that can be developed upon the client s request Signum Dermalogix is the solution to streamline and optimize your R&D screening, management, and/or customized research at a reduced cost. We provide superior service to differentiate your compound/ingredient. We will provide the scientific excellence and quality your product deserves. Contact us today. 1

3 STANDARD ASSAY SYSTEMS Skin Inflammation Skin Aging Pollutant Toxicity Anti Microbial Anti Oxidant Skin Whitening Skin Tanning Wound Healing Metabolism Safety Testing 8 SIGNUM CUSTOMIZED SCREENING SERVICES 9 SIGNUM COMPOUND PROFILE SCREENING SERVICES 10 SIGNUM ANALYTICAL SERVICES SERVICES 11 2

4 SKIN INFLAMMATION Catalog # Cell line Stimulus/Inducer Endpoint Method eference SKI 001 Epidermis (EpiDerm) TPA/PMA IL 8 IHC Clobetasol SKI 002 Epidermis (EpiDerm) TPA/PMA PGE 2 IHC Indomethacin SKI 003 Epidermis (EpiDerm) UVB IL 8 IHC Clobetasol SKI 004 Epidermis (EpiDerm) UVB Sunburn Cells IHC Vitamin C SKI 005 Epidermis (EpiDerm) UVB DNA pyrimidine dimers IHC Vitamin C SKI 006 Epidermis (EpiDerm) UVB/UVA Lipid Peroxide IHC Vitamin C SKI 007 Keratinocytes (NHEK) TPA/PMA IL 8 IF Clobetasol SKI 008 Keratinocytes (NHEK) TPA/PMA PGE 2 IF Indomethacin SKI 009 Keratinocytes (NHEK) UVB IL 8 IF Clobetasol SKI 010 Keratinocytes (NHEK) UVB Sunburn Cells IHC Vitamin C SKI 011 Keratinocytes (NHEK) UVB DNA pyrimidine dimers IHC Vitamin C SKI 012 Keratinocytes (NHEK) H2O2 phospho JNK IHC SP SKI 013 Keratinocytes (NHEK) UVB phospho JNK IHC Vitamin D3 SKI 014 Keratinocytes (NHEK) IL 1alpha IL 8 IF Clobetasol SKI 015 Keratinocytes (NHEK) ATP gs IL 6 IF Clobetasol SKI 016 Peripheral Blood Mononuclear Cells (PBMC) CD3/CD28 IL 4 ELISA TGF beta SKI 017 Peripheral Blood Mononuclear Cells (PBMC) CD3/CD28 IL 17 ELISA TGF beta SKI 018 Human Dermal Microvascular Endothelial Cells (HDMEC) ATP gs MCP 1 ELISA Clobetasol 3

5 SKI 019 SKI 020 Human Dermal Microvascular Endothelial Cells (HDMEC) anti microbial peptides(ll 37) IL 6 ELISA Clobetasol Keratinocytes (NHEK) P. acnes IL 8 ELISA Dexamethasone SKIN AGING SKA 001 Keratinocytes (NHEK) Cell Confluence Keratin 1 WB Adapalene SKA 002 Keratinocytes (NHEK) Cell Confluence Keratin 1 WB Adapalene SKA 003 Keratinocytes (NHEK) Cell Confluence Filaggrin WB Adapalene SKA 004 Keratinocytes (NHEK) Cell Confluence Loricrin WB Adapalene SKA 005 Fibroblasts (HDF a) Basal Collagen ELISA, WB Vitamin C SKA 006 Fibroblasts (HDF a) Basal Elastin ELISA, WB Vitamin C SKA 007 Fibroblasts (HDF a) UVA Collagen ELISA, WB Vitamin C SKA 008 Fibroblasts (HDF a) UVA Elastin ELISA, WB Vitamin C SKA 009 Fibroblasts (HDF a) H2O2 Collagen ELISA, WB Vitamin C SKA 010 Fibroblasts (HDF a) H2O2 Elastin ELISA, WB Vitamin C SKA 011 Fibroblasts (HDF a) Basal Fibronectin ELISA, WB TGF beta SKA 012 Fibroblasts (HDF a) Basal Laminin ELISA, WB TGF beta SKA 013 Fibroblasts (HDF a) UVA Fibronectin ELISA, WB TGF beta SKA 014 Fibroblasts (HDF a) UVA Laminin ELISA, WB TGF beta SKA 015 Fibroblasts (HDF a) H2O2 Fibronectin ELISA, WB TGF beta SKA 016 Fibroblasts (HDF a) H2O2 Laminin ELISA, WB TGF beta SKA 017 Fibroblasts (HDF a) Basal Hyaluronic acid ELISA, WB TGF beta SKA 018 Fibroblasts (HDF a) Basal Hyaluronic acid ELISA, WB TGF beta SKA 019 Fibroblasts (HDF a) UVA Hyaluronic acid ELISA, WB TGF beta 4

6 SKA 020 SKA 021 SKA 022 SKA 023 SKA 024 SKA 025 Fibroblasts (HDF a) H2O2 Hyaluronic acid ELISA, WB TGF beta Fibroblasts (HDF a) Basal MMP 1 ELISA Dexamethasone Fibroblasts (HDF a) UVA MMP 1 ELISA Dexamethasone Fibroblasts (HDF a) H2O2 MMP 1 ELISA Dexamethasone Cell Fibroblasts (HDF a) in vitro wound Migration IF FCS Keratin Epidermis (EpiDerm) Basal 1,10 IHC Adapalene POLLUTANT TOXICITY TOX 001 Epidermis (EpiDerm) Cadmium/Nickel MTT Reduction Colorimetric EGF TOX 002 Keratinocytes (NHEK) Cadmium/Nickel MTT Reduction Colorimetric EGF TOX 003 Fibroblasts (HDF a) Cadmium/Nickel MTT Reduction Colorimetric TGF beta TOX 004 Human Dermal Microvascular Endothelial Cells (HDMEC) Cadmium/Nickel MTT Reduction Colorimetric Glucose TOX 005 Human Dermal Microvascular Endothelial Cells (HDMEC) Nickel IL 6 ELISA Clobetasol TOX 006 Human Dermal Microvascular Endothelial Cells (HDMEC) Nickel IL 8 ELISA Clobetasol 5

7 ANTI MICROBIAL AMIC 001 Keratinocytes (NHEK) Basal LL 37 IHC AMIC 002 Keratinocytes (NHEK) LPS IL 8 IF Doxycycline AMIC 003 Human Umbilical Vein Endothelial Cells (HUVEC) LPS IL 8 IF Doxycycline AMIC 004 P. acnes Basal Cell death OD Doxycycline AMIC 005 S. epidermidis Basal Cell death OD Doxycycline AMIC 006 E. coli Basal Cell death OD Doxycycline ANTI OXIDANT AOX 001 Human Neutrophils HL 60 fmlp/pma Cell Oxidative Burst Colorimetric Vitamin E AOX 002 N/A Basal TBARS Colorimetric Trolox AOX 003 N/A Basal LDL Oxidation Colorimetric Vitamin C AOX 004 N/A Basal Microsome Oxidation Colorimetric Vitamin C 6

8 SKIN WHITENING SKW 001 Epidermis (MelanoDerm) Basal Melanin Colorimetric Kojic Acid/ Iklen Rucinol SKW 002 Epidermal Melanocytes Basal Melanin Colorimetric Kojic Acid/ Iklen Rucinol SKIN TANNING SKT 001 Epidermis (MelanoDerm) Basal Melanin Colorimetric IBMX SKT 002 Epidermal Melanocytes Basal Melanin Colorimetric IBMX WOUND HEALING WH 001 Keratinocytes (NHEK) Cell Confluence Keratin 1 WB Adapalene WH 002 Keratinocytes (NHEK) in vitro wound Cell Migration IF EGF 7

9 METABOLISM MET 001 Keratinocytes (NHEK) Basal Mitochondrial localization IF FCS MET 002 Epidermis (EpiDerm) Basal Mitochondrial localization IF FCS MET 003 Keratinocytes (NHEK) Basal Ki66, Epsin 3 IF EGF MET 004 Epidermis (EpiDerm) Basal Ki66, Epsin 3 IF EGF SAFETY TESTING SAF 001 Epidermis (EpiDerm) Basal Skin Irritation Colorimetric SDS SAF 002 Skin (EpiDerm FT) UVA Photodamage IHC Chlorpromazine SAF 003 Skin (EpiDerm FT) UVB Photodamage IHC Sun Screen SAF 004 Skin (EpiDerm FT) IR A Photodamage IHC Vitamin C SAF 005 Epidermis (EpiOcular) Basal Animal free Draize Test Colorimetric SDS SAF 006 Mouse BALB/3T3 Fibroblasts UVA Cell Viability Neutral Red Assay Chlorpromazine SAF 007 Keratinocytes (NHEK) Basal Cell Viability MTS Assay SLS SAF 008 N/A Daylight: D65 Chemical Degradation HPLC quinine monohydrochloride dihydrate 8

10 Standard dermatological tests while effective in many respects do not always provide the best path forward in characterizing the activity of your molecule or product. Given our past experience in working to develop and commercialize cosmetics, extracts, pharmaceuticals and nutraceuticals, we possess innovative capabilities in protocol design, assay development and claim support testing. With the support of our knowledgeable project management team, we can demonstrate the activity of your molecule or product and tailor the appropriate assay systems to produce exciting scientific data to deliver your study objectives and support your product s brand goals. Contact us today to select the tests that best fit the vision and criteria of your compound. Together, we will develop the appropriate dermatology assay project that will provide you with robust scientific reasoning, results and claims for your product. Design Characterization Optimization Analysis 9

11 Profile 001 Multiple Several Multiple Several Assay specific To identify and/or characterize molecules that have beneficial effects on skin cells, Signum Dermalogix has developed a broad based screen to test your compound, ingredient or extract. This initial screening strategy is employed to assess a small molecule s activity and comparing it to several standard reference compounds that are broad acting in their activities, since the dermal inflammatory response is realized through interplay between a numbers of different cell types: keratinocytes, endothelial cells, fibroblasts and various leukocytes. Initial profiling efforts test the effects of compounds in several commonly accepted cellular models of inflammation utilizing a number of these cell types. In addition to measuring inhibitory effects of various markers of inflammation, testing for antimicrobial activity and evaluating effects on keratinocyte differentiation is also performed. Altogether this initial characterization will determine your compound of interest s activities over a broad number of cellular systems and molecular endpoints. This type of profiling is especially valuable in identifying potentially new or additional claims for your molecule that may have previously been missed or during early stage discovery. This screening program will*: Identify the skin and immune cell types affected by the compound Discover the specific molecular processes that are modulated by the compound Determine if the small molecule has anti bacterial activity versus skin flora Determine activity of each molecule in regulating cellular processes & microbial growth Discover the viability of several skin & immune cell types in the presence of the compound Determine if the compound has anti oxidant properties *Signum Dermalogix can customize screening to your own specification and target(s) of interest. Assays may be swapped if specific tests listed in our standard assay system are of greater interest for studying your molecule. 10

12 Skin layer SPEN 001 Pig Skin (ex vivo) Basal levels Franz Cell Vehicle Once a compound or product is found to be effective in vitro, the next steps in development for skin care call for the compound to be formulated to assess it dermal penetration profile, stability and further safety testing. In addition to custom assay development and characterizing the activity and safety profile of your compound or product, Signum Dermalogix provides analytical and testing services including: Analytical support (HPLC, LC MS, NMR) Method development Stability assessment Radioisotope labeling Franz cell dermal penetration studies Pre formulation assessment In vivo safety assessment We maintain an NRC license to use radioisotopes and have an Institutional Animal Care and Use Committee (IACUC) in place for SOP approval and compliance with animal safety testing regulations. Early development and precise strategy can reduce costs and accelerate timelines. Contact us to learn more about our services and where Signum Dermalogix can bring maximum benefit to your development program and product. 11

13 The Science of Staying Healthy Eduardo Perez, PhD Chief Scientific Officer ext

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