Sytenol®A Skin protection Sytenol®A for Skin Protection Extended life-span, more spare time and excessive exposure to UV radiation from sunlight or tanning devices, especially in the Western population, has resulted in an ever increasing demand to protect human skin against the detrimental effects of UV-exposure. It is well-known that UV light acts directly on nucleic acids or indirectly via reactive oxygen species, resulting in altered redox balance, activation of repair processes, and altered gene expression in skin. Sunscreens - the current gold standard of photo-protection - are useful, but their skin protection ability is inadequate against UV-induced skin damage. The most severe consequence of photo-damage is skin cancer. Less severe photo-aging changes result in wrinkling, scaling, dryness, and uneven pigmentation consisting of hyper- and hypo-pigmentation. Sytenol® A in combination with photostable broad-spectrum sunscreen provides a solution to skin protection and treatment. Product Information Trade Name Sytenol® A INCI Name Bakuchiol CAS # 10309-37-2 ELINCS N/A Harmonized Tariff # 29071190 Appearance Yellow viscous liquid Assay (GC) 95% (w/w) min Bakuchiol content Solubility Highly miscible with a wide range of hydrophobic emmolients and solubilizers Suggested use 0.25 to 1% Storage Store in original, sealed container at +10 to +30 0C; Avoid light & heat Patent status Covered by multiple US and world-wide patents Safety Data: REACH-ready safety profile available upon request q q q q q No skin irritation & No skin sensitization (HRIPT) at 2 and 5% dilution in corn oil No eye irritation (In-vitro; alternate to Draize methodology) Non-genotoxic in the in-vitro micronucleus assay in human lymphocytes Non-mutagenic based on TA98 in Salmonella typhimurium Reverse Mutation Assay Passes aquatic toxicity tests (short-term, Zebra fish & Daphnia magna); Fresh water single cell green alga & biodegradability Regulatory Status: USA: Allowed for personal care applications Europe: Up to 1 Ton annual use requires no REACH registration Rest of the World: Allowed for use in most Asian, South American and African countries Consult your regulatory authority for any restrictions this product may have for use in your country Sytenol® A Protects Skin from UV-induced Damage Erythema: Cause & Consequences: Erythema, the most familiar manifestation of UV radiation exposure, occurs in a biphasic manner. UVA mediates the early part of this reaction, known as immediate pigment darkening (IPD) and lasts for about half-hour. Delayed erythema, a function primarily of UV-B dosages, begins 2-8 hours after exposure and reaches a maximum in 24-36 hours, with erythema, pruritius, and pain in the sun-exposed areas. A dose of UVB radiation sufficient to induce erythema in human skin results in the formation of about 20 photoproducts per 106 nucleotides. qCyclooxygenase (COX)-dependent prostaglandin E2 (PGE2) is a mediator of UVR-induced erythema qPhospholipase A2 (PLA2) is a rate limiting step in the generation of leukotrienes and prostaglandins. PLA2 synthesis occurs only when skin is exposed to UV doses that are sufficient to cause erythema qDNA may be a chromophore for erythema. Pyrimidine dimerization in transcriptionally active DNA is a key contributor to UV-induced erythema and inflammation qNitric oxide (NO) is also a contributor to the UV erythema response. NO is produced in the skin by NO synthase that can combine with superoxide to form peroxynitrile, a highly reactive oxidant and mediator of erythema and cause of tissue injury. Protocol: Human Clinical Study Sytenol®A Sytenol®A SUMMARY Treated & irradiated site with 1% Sytenol® A lotion: Showed practically no changes in L, a and b-values and ITA0 (average of 10 subjects) n Conclusion: Protection of skin against UV-induced erythema/inflammation Untreated & irradiated site: Showed statistically significant differences in L-, a- and b-values and ITA0 (average of 10 subjects) n Conclusion: No protection; Significant skin erythema/inflammation under UV-exposure *Statistically significant; p < 0.001 Sytenol® A Provides Skin Protection by Quenching Radicals & Non-radicals and by Up-regulating Antioxidant Defense System Sytenol® A is a Broad-Spectrum Antioxidant Sytenol® A Up-regulates Antioxidant Defence Genes Antioxidant Defense Gene Expression Glutathione peroxidase 3 precursor/ Extracellular glutathione peroxidase(GPX3): Protect organism from oxidative damage. Reduce lipid hydroperoxides --> alcohols and hydrogen peroxide --> water Glutathione S-transferase theta -1 (GSTT1): Involved in the detoxification of endogenous compounds, such as peroxidised lipids, as well as the metabolism of xenobiotics Glutathione S-transferase P 1(GSTP1): Same as above NAD(P)H dehydrogenase [quinone] (NQO1):This protein’s enzymatic activity prevents the one electron reduction of quinones that results in the production of radical species. Fold Change vs Control Sytenol® A Protects Skin via Multi-factorial Oxidative Pathways Literature Data q Inhibits mitochondrial lipid peroxidation [H Haraguchi et al., Planta Med, 66(6):569-571, 2000] q Protects mitochondrial respiratory enzyme activities against both NADPH-dependent and dihydroxyfumarate-induced peroxidation injury [H Haraguchi et al., Planta Med, 66(6):569571, 2000] q Prevents NADH-dependent and ascorbate-induced mitochondrial lipid peroxidation; Protects human red blood cells against oxidative haemolysis; Protects biological membranes against various oxidative stresses [H Haraguchi et al., Phytother Res, 16(6):539-44, 2002] q Inhibits the expression of inducible nitric oxide synthase gene via the inactivation of nuclear transcription factor-kappaB in RAW 264.7 macrophages [HO Pae et al., Intl Immunopharmacol,1 (9-10):1849-55, 2001] q Inhibits lipid peroxidation induced by Fenton-reaction, [S Adhikari et al., Chem Res Toxicol, 16:1062-1069, 2003] q Induces caspase-3-dependent apoptosis through the activation of JNK, followed by Bax translocation into mitochondria in rat liver myofibroblasts [EJ Park et al., Eur J Pharmacol, 559 (2-3):115-123, 2007] Sytenol® A Protects Skin by Inhibiting Glucose Oxidase Activity % Reduction in Glucose Oxidase Activity Assayed in vitro with the Amplex® Red Glucose/Glucose Oxidase Assay Kit (Invitrogen, cat. #A22189) q nGlucose oxidase converts glucose to glucono-d-lactone with the release of H2O2 + Iron --> Reactive Oxygen Species --> Oxidative stress to skin (Fenton reaction) nH2O2 Sytenol® A Provides Skin Protection by Down-regulating Pro-inflammatory Genes and Enzymes Sytenol® A Down-regulates Pro-inflammatory Genes (DNA microarray study) Pro-inflammatory gene expression Cyclooxygenase-1 (Prostaglandin G/H synthase precursor)(PTGS1/ COX-1): Prostaglandin biosynthesis; Acts both as a dioxygenase and as a peroxidase Phospholipase A-2-activating protein (PLAA): Eicosanoid synthesis, release of neutrophil lysosomal enzymes and superoxide and on RBC hemolysis Cytosolic phopholipase A2 (PLA2G4A): Catalyzes hydrolysis of membrane phospholipids to release arachidonic acid which is subsequently metabolized into eicosanoids Prostaglandin E2 receptor EP2 subtype (PTGER2): Inflammatory reaction via the EP2 receptor through its regulation of TNF-alpha and IL-6 Prostaglandin E2 receptor EP4 subtype (PTGER4): Inflammatory reaction via the EP4 receptor through its regulation of TNF-alpha and IL-6 Prostaglandin dehydrogenase 1(HPGD): Degrades PGF2a and PGE2 (cause inflammation) and converts to hydroxy metabolites (anti-inflammatory) Fold change vs Control Sytenol® A Inhibits Pro-inflammatory Enzymes Literature Data Phopholipase A2 inhibitor: Weak inhibitor of Phospholipase A2, but dose-dependently inhibits Leukotriens B4 and Thromboxane B2; Topical administration inhibited TPA-induced ear edema and myeloperoxidase activity and also reduced PGE2 content in arachidonic acid acid-induced response [ML Fernandiz et al.,J Pharm Pharmacol, 48:975–980, 1996] q iNitric Oxide Synthase (iNOS) inhibitor: Inhibits expression of iNOS mRNA through the inactivation of transcription factor -kB [HO Pae et al., Intern Immunopharmacol, 1(9-10), 1849-1855, 2001 q New Sytheon Data Cyclooxygenase inhibitor: ~40% reduction in COX-2 inhibition with 50 µg/ml Sytenol A; Assayed in-vitro using a kit from Cayman Chemical; Determined COX-2 activity at 590 nm with BioRad 3550-UV microplate spectrophotometer q Sytenol®A Provides Skin Protection by Inhibiting Elastase Gene Expression Elastase breaks down elastin, an elastic fiber that, together with Collagen determines the mechanical properties of connective tissue. Sytenol®A also has a moderate inhibitory activity against Collagenase (MMP-1), which is responsible for degrading Collagen I. The major component of extracellular matrix is Collagen, a support system for the survival of each and every cell in the body. Sytenol®A up-regulates Elastase-specific inhibitory genes [DNA micro-array study] Gene Gene Description PI3 Elastase-specific inhibitor SERPINB1 Leukocyte elastase inhibitor Function Suppresses degradation of connective tissue components, such as elastin, and collagen. Suppresses the development of solar elastosis and skin tumors Fold Change Vs. Control Sytenol® A Retinol +2.7 +2.5 +6.1 +3.8 In-Vitro Elastase Inhibitory Activity: Sytenol®A vs. Retinol Sytenol® A has a reverse dose-dependency in inhibiting elastase activity whereas Retinol has a stimulatory activity Assayed using Calbiochem human neutrophil elastase (Cat # 324681) % Reduction in Elastase Activity & its substrate (Elastase substrate VIII). Positive control – El III (Calbiochem Catalog # 324745) & negative control water Elastase inhibitory activity was determined by quantifying the absorbance of chromophoric reaction product at 410 nm using BioRad microplate reader Sytenol®A Provides Skin Protection by Inhibiting Hypoxia Inducible Factor 1 (HIF-1) [J Biol Chem, 282(22):16413-16422, 2007] q q q q HIF-1 is a stress inducible transcription factor; triggers during hypoxia such as, in tumor and in ischemic cardiovascular disease and also in normoxia such as, UV-induced Reactive Oxygen Species generation in keratinocytes HIF-1 regulates different genes involved in cell survival, apoptosis, cell motility, cytosketal structure, cell adhesion and energy metabolim Pro-stimulatory effect on HIF-1 causes photocarcinogenesis HIF-1 Inhibitory activity of Bakuchiol: IC50 6.1 µM [Tetrahedron Letters, 48(50):8861-8864, 2007] Sytenol®A Protects the Skin via Multi-factorial Pathways Sytenol® A clearly shows promise as a new agent that can complement and enhance the photo-protective effect of currently available sunscreens. In-vitro and studies in humans, Sytenol® A ameliorates many of the adverse effects of sun-induced skin damage triggered by multiple pathways. It is a potent broad-spectrum antioxidant, is an effective inhibitor of a wide range of pro-inflammatory genes and enzymes and has a matrix-degrading metalloprotease inhibitory activity. Sytenol® A has an excellent safety profile and is a non primary skin irritant and a non primary skin sensitizer as demonstrated by Human RepeatInsult Patch Test [HRIPT] studies. Topical formulations that include Sytenol® A are likely to lead further improvements in the way we protect our skin from overexposure to sun. USA: 315 Wootton Street, Boonton, NJ 07005 www.sytheonltd.com - [email protected] Disclaimer The information given and the recommendations made herein are based on our research and literature search and are believed to be accurate but no guarantee of their accuracy is made. This information is intended to be helpful, but no warranty is expressed or implied as to the results obtained from use in the formulation, procedure or products suggested herein. Neither is any permission or recommendation to practice any invention covered by patent either expressed or implied. Tel.: +1 973.988.1075 FRANCE: 1ère avenue, 1ère rue, BP 383 06514 Carros Cedex www.sytheonltd.com - [email protected] 2011 - www.growitgroup.com Tel.: +33 (0)4.92.08.52.42