Arch Personal L.P. Soy Zymbiozome Fermentum (Patent Pending) Code #136430 Assigned INCI Designation: Water (and) Glycine Max (Soybean) Symbiosome Extract Product Information Drawing on the resources of Arch Personal product innovation groups, we have developed an entirely new class of patent-pending cosmetic ingredients based on root nodules derived from Soybeans (Glycine max). Glycine max symbiosome extract is isolated from symbiosome lysates harvested in the early spring planting of soybeans when the root nodules are at their most active and potent state. The root nodules are harvested, cleaned, macerated and the unique active components are extracted by aqueous extraction technologies core to Arch s manufacturing expertise. Working in collaboration with academic researchers exploring the unique properties of leghemoglobin, we standardized these products in leghemoglobin content. Because of the extraordinary relationship between plantbased leghemoglobin, and human based hemoglobin and cytoglobin, we felt that this particular extract might have nitric oxide binding capabilities comparable to human hemoglobin. 1 In the human body, hemoglobin is partly responsible for converting nitric oxide into less reactive chemical species such as nitrates. 2 The role of nitric oxide in the human body is both beneficial and potentially damaging if the concentrations of this gaseous agent exceed the body s ability to remove it. Nitric oxide in the skin is felt to be part of the biochemical cascade that is responsible for the formation of redness during sunburn and irritation. 3 Key Product Attributes A globin protein, leghemoglobin, as active component Anti-inflammatory agent Free radical scavenger Preventative mechanism against photo-aging 11/24/04 v1.8
Arch Personal L.P. Product Soy Zymbiozome Fermentum, standardized to leghemoglobin, offers unique nitric oxide scavenging properties along with a powerful package of additional antioxidants and skin hydrating agents that further enhance the functionality of this product as a preventative ingredient to combat photoaging of the skin. 5-7 Figure 1. Realms of Prevention and Cure. Soy Zymbiozome Fermentum clearly falls under prevention; mitigating the visible signs of aging. Re al m of Pre ve nti on Realm of Prevention Re al m of Cu re Realm of Cure Time Minutes to Hours Hours to Days Absorbed UV Radiation and O2 Generation of free radicals (Super Oxide, Hydroxide, Nitric Oxide) Anti - oxidants Vitamins Stimulation of biochemical cascades (IL - 1 α, TNF - α, PGE2, NF - κβ, MSH, etc ) Anti - inflammatories Days to Weeks Sub - clinical damage ( Erythemia, Microscarring, Matrix degradation) Weeks to Years Visible damage (Sunburn, Wrinkles, Photoaging ) Prevention Sunscreens Retinol, AHA s, BHAs
Arch Personal L.P. Technical In vitro Testing Full-Thickness Human Tissue Model Studies Human full-thickness tissue models provide an improved means for determining the effectiveness of test products to influence skin physiology and begin to provide some idea of the ability of an active to penetrate the stratum corneum. 8 MatTek Epi-Derm EFT-200 was employed as a test substrate to examine the influence of 1% Soy Zymbiozome Fermentum on a number of key enzymatic markers responsible for sub-clinical erythema. Ultraviolet radiation was employed as an activator for the enzymes and as a stimulus for nitric oxide production. The key enzymatic markers for irritation included IL-1a, TNF-a, and PGE 2. In addition, a key enzyme responsible for skin collagen and elastin breakdown, active MMP-1, was examined. The tissue model also allowed for examination of any cytotoxic effects due to the presence of the test material and for the examination of the ability of the test product to control nitric oxide in human skin tissue. Results of the entire test are compiled in Table 1. Table 1. Effect of UV radiation and 1% Soy Zymbiozome Fermentum on full-thickness tissue including effects on enzyme response, tissue viability and nitric oxide production. An arrow pointing up indicates a statistically significant increase in the measured activity and an arrow pointing down indicates a reduction in measured activity. for the UV irradiated tissue is compared against un-irradiated controls and data for 1% Soy Zymbiozome Fermentum is compared against data from the irradiated tissue. Marker Testing Method UV UV+ Soy Tissue Viability MTT No Effect NO Griess Reaction Interleukin-1α ELISA No Effect Prostaglandin E 2 ELISA Tumor Necrosis ELISA Factor-α Active Matrix Metalloproteinase-1 ELISA Results The data indicates that irradiation of the tissue with ultraviolet light causes increases in production of nitric oxide, reduces cell viability and increase production of both the irritation cascade enzymes and the collagenase enzyme activity. Application of 1% Soy Zymbiozome Fermentum controls the production of two of the three enzymatic markers for erythema, influences the marker for collagenase enzyme and reduces the production of nitric oxide due to UV exposure.
Arch Personal L.P. Product In vivo Testing An in vivo study was done to demonstrate the anti-irritation effect of Soy Zymbiozome Fermentum after UV irradiation. Sub-clinical erythema was measured using cutaneous microcirculation via Laser Doppler, a test method suggested to be the most sensitive for measuring sub-clinical changes in the skin. 9 The test protocol included the following applications: untreated site, placebo (water and gelling agent), preventative zone application prior to UV exposure (1% Soy Zymbiozome Fermentum), curative zone application immediately after UV exposure, curative zone application 2 hours post exposure. Test participants were exposed to 1.0 MED broad-spectrum ultraviolet radiation treatment. An increase in microcirculation is correlated to an increase in sub-clinical erythema, whereas a decrease in microcirculation is indicative of a decrease in sub-clinical erythema. Figure 2 presents the test data as the difference between hour 24 measurements and hour 12 measurements, providing a snapshot of the effect of the test product to reduce redness during the 12-hour natural healing process. Figure 2. Anti-inflammatory effect of Soy Zymbiozome Fermentum as both a preventative and curative application. Time 24 (Hours) -Time 12 (Hours) Difference in Microcirculation Value (T24-T12) 20 10 0-10 -20-30 -40-50 Non-Treated Zone Placebo Treated Preventative Zone Treatment Curative Zone Curative Zone (2H)
Arch Personal L.P. Discussion The in vitro and in vivo test results outlined above support the theory regarding the mechanism of activity of Soy Zymbiozome Fermentum and the principal active component, leghemoglobin. From the results of the testing data it appears that leghemoglobin binds NO as it is formed by UV irradiation. If the active is applied before free radical nitric oxide is formed, the leghemoglobin effectively binds it and minimizes increases in sub-clinical erythema. If the active is applied just after application of the radiation, the reductions in sub-clinical erythema are still significant, but not as large as those seen if the product is present during irradiation. Application of Soy Zymbiozome Fermentum several hours after irradiation reduces the effectiveness of the product. This data supports the conclusion that the active ingredient in Soy Zymbiozome Fermentum, i.e., leghemoglobin, is a powerful free radical scavenger, but that it must be present at the time of the production of the free radicals in the skin or the natural defense mechanisms of the skin begin to remove the excess free radicals. Conclusion Soy Zymbiozome Fermentum minimizes the free radical induced irritation cascade process resulting from UV exposure of the skin. Reduction in production of free radical nitric oxide helps to lower the resulting redness (sub-clinical erythema) associated with high UV exposure on the skin. Reductions in sub-clinical erythema can help to prevent the long-term adverse effects of solar radiation on the skin including photoaging, wrinkles and sagging. Cosmetic Applications Sun o Preventative and curative benefits o Free radical quenching, redness reduction Daily wear lotions o Prevents first signs of aging Decorative Cosmetics o Treatment products prevention Superior Anti-Aging Technology Toxicological/Safety Review Epi-Ocular Non-Irritating Epi-Derm Non-Irritating RIPT Non-Irritating AMES Non-Mutagenic Soy Allergens None Endotoxins None Trace Metals Lead Pb (ppm) < 1 Arsenic As (ppm) < 1 Mercury Hg (ppm) < 1
Arch Personal L.P. References 1. Pickrell, J. Globin Family Grows: Blood-protein relative is in all tissues. Sci News. 2002, 161, 230. 2. Yoshida, K. et. al., Metabolic fate of nitric oxide Int Arch. Occup. Environ. Health 1980, 46, 71-77. 3. Bruch-Gerharz, D. et. al., Nitric oxide in human skin: current status and future prospects J. Invest. Dermatol. 1998, 110, 1-7. 4. Wilson, E.K. New NO Directions Chem. Eng. News 2004, 82, 39-44 5. Iturbe-Ormaetxe, I. et. al., The antioxidants of legume nodule mitochondria Mol. Plant Microbe Interact. 2001, 14, 1189-1196. 6. Okker, R. J. H. et. al., Flavonoids in the rhizobium-legume symbiosis Prog. Clin. Biol. Res. 1988, 280, 71-76. 7. Karg, G. et. al., Protective role of natural antioxidants Cosmet. Toilet. 1987, 102, 37-51. 8. Ponec, M. et al., Characterization of Reconstructed Skin Models. Skin Pharmacol. Appl. Skin Physiol 2002, 15, 4-17. 9. Zuang, V. et al., Detection of skin irritation potential of cosmetics by non-invasive measurements, Skin Pharmacol Appl. Skin Physiol 2000, 13, 358-371.
Arch Personal L.P. Product DAILY WEAR SPF-12 SUNSCREEN WITH SOY ZYMBIOZOME ST-48 Soy Zymbiozome provides total skin protection and age prevention in one ingredient! This powerful free radical scavenger prevents skin redness due to erythema and is useful in daily wear products to provide continual protection. Gel Base BSM-PE provides a powdery, silky after-feel. Liqiuiwaxes DIADD and PolyEFA are excellent pigment dispersants for the physical sunscreen and add a cushiony feel to this daily wear product. Ingredient INCI Nomenclature Supplier % Phase A Water Water - q.s. Glycerin Glycerin - 2.00 Sodium Chloride Sodium Chloride - 1.00 Germaben II Propylene Glycol & Diazolindinyl Urea & ISP 1.00 Methylparaben & Propylparaben Water & Glycine Max (Soybean) Symbiosome Extract Soy Zymbiozome Arch Personal Fermemtun 1.00 Phase B Dow Corning 345 Cyclomethicone Dow Corning 7.50 DC 200/100 cst Dimethicone Dow Corning 1.00 Escalol 557 Ethylhexyl Methoxycinnamate ISP 7.50 Liquiwax DIADD* Dioctyldodecyl Dodecanedioate Arch Personal 1.00 Gel Base BSM-PE Cyclomethicone & Dimethicone & Phenyl Arch Personal Trimethicone & Polyethylene 3.00 Liquiwax PolyEFA* Octyldodecyl PPG-3 Myristyl Ether Dimer Arch Personal Dilinoeate 5.00 Z-Cote Zinc Oxide BASF 5.00 Abil EM-90 Polyglyceryl-4 Isostearate & Cetyl Degussa Care PEG/PPG-10/1 Dimethicone & Hexyl Specialties Laurate 3.00 Fragrance Fragrance - q.s * Manufactured by Innovachem and distributed by Arch Personal. Specifications: ph: N/A Viscosity: 60,000 80,000 cps (TE, 10RPM @25 C) Procedure: 1. Combine Phase A ingredients and mix until uniform. 2. Combine Phase B ingredients and mix until uniform. 3. Slowly add Phase A to Phase B with prop mixing. Mix until homogeneous.
Arch Personal L.P. RADICAL SPRINKLE BT-13 This cold-mix formulation uses a very stable Sub Micron Emulsion Concentrate (SMEC Concentrate # 1) consisting of particles in the region of 250 nm. This makes very low surfactant levels possible and, because of deeper skin penetration, really moisturizes the skin. Soy Zymbiozome Fermentum offers unique nitric oxide scavenging properties along with additional antioxidants, skin hydrating agents and moisturizers, resulting in a fragranced body spray that leaves the skin refreshed, moisturized and silky smooth. Ingredient INCI Nomenclature Supplier % Demineralized Water Demineralized Water q.s. SMEC Concentrate #1 Trimethylolpropane Tricaprylate/Tricaprate & Glycerin & Cetearyl Alcohol & Ceteareth-20 & Glyceryl Stearate & PEG-100 Stearate & Steareth-2 & Dimethicone & Ceteth-24 & Choleth-24 & Phospholipids & Propylene Glycol Water & Glycine Max (Soybean) Arch Personal Soy Zymbiozome Arch Personal Fermentum Symbiosome Extract 0.1 Fragrance Fragrance 1.0 Polysorbate 20 Polysorbate 20 1.0 50.0 Procedure: 1. Add the SMEC Concentrate to the water and mix well. 2. Add the Soy Zymbiozome Fermentum while stirring slowly. 3. Pre-mix the fragrance and Polysorbate 20 and add to the batch. Mix well.