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PERSONAL CARE Vincience™ Biofunctionals Chronogen™ Description Innovative anti-aging tetrapeptide, designed using advanced molecular biology and inspired by epigenetic science, to boost the expression of clock genes The circadian rhythm is an important regulatory function that skin cells need to regenerate and revitalize for a fresher appearance. This internal clock serves to optimize repair activities around the day-night cycle of the earth’s rotation and is synchronized with other cells and organs operating under the same schedule. For many years, time-keeping functions were thought to be exclusively centralized in the brain within the suprachiasmatic nucleus. Although this master clock plays an important role, recent research has shown that cells can independently “tell time” through a very precise molecular mechanism involving the expression and regulation of circadian genes (Clock, Per1, Bmal1, Tim and Cry). These clock genes give tempo to cells, orchestrating and regulating more than 20% of gene expressions. Factors such as UV radiation (even in low doses) can decrease the expression of these clock genes and “desynchronize” the cells’ circadian rhythm. Benefits • Enhances expression of CLOCK, BMAL1 and PER1, in-vitro and ex-vivo • Helps preserve skin from UV damage during the day • In-vitro, helps better DNA repair The bio-engineered peptide contained in Chronogen has been designed to improve the expression of the proteins involved in the clock mechanism and represents a major breakthrough strategy in cosmetic science. Cosmetic Applications Because consumers often apply cosmetic products around a day-night regimen, the use of formulations aiming the circadian rhythms may help further optimize cellular protection and renewal. In-vitro results show that Chronogen specifically boosts cellular natural defenses against UV damage (important during the day) and helps better natural DNA repair (important at night). • Day time formulations to help boost skin’s natural defenses against UV • Night time formulations to boost the skin’s repair process • Innovative anti-aging formulations based on epigenetic science • Formulations to help optimize the protective and regenerative cycle of skin I Introduction For many years, the consensus was that time-keeping functions were highly centralized in a master clock located in the suprachiasmatic nucleus within the brain. More recently, discoveries have shown that each cell exhibits a deeply embedded self-sustained circadian rhythm used to govern gene expression. This mechanism has become a major focus in biology and medical research. With Chronogen, ISP is introducing to the personal care market an advanced technology, inspired by epigenetic science, designed to help consumers optimize skin regeneration and the natural defenses against UV damage. Circadian Regulators: A Cellular Metronome The skin cells’ defense and repair is orchestrated by each cell’s own internal “clocks”. Preserving optimal expression of these clock genes is a challenge because even low UV exposure can knockdown these regulating clock genes for more than 24 hours. The time-keeping mechanism is modeled as a network of transcriptional and post-translational feedback loops synchronized on the day-night cycles. It involves several genes including Clock, Per1, Bmal1, Tim and Cry. They help to regulate the cycle in which cells turn on and off repair functions, ensuring to coordinate these functions with UV exposure. Hence, targeting clock genes represents a major breakthrough in anti-aging strategy in cosmetic science. CLOCK PER1 + 41% + 52% Control Chronogen (1%) Quantification Cell Culture: Human skin biopsy Product Application: Chronogen (1%), twice a day Application Time: 62 hours Evaluation: Immunofluorescence staining in green of circadian proteins (CLOCK or PER1) Quantification: Image analysis In-vitro and ex-vivo tests suggest that Chronogen can boost the expression of circadian gene proteins: CLOCK, BMAL1 and PER1. Chronogen provides an innovative anti-aging breakthrough strategy by aiming for optimal expression of the internal clock gene proteins. DNA Repair and Protection DNA is constantly submitted to damaging stresses (UV rays and free radicals). In particular, when UV exposure is the highest during the day, DNA strands must be configured in a way to ensure maximum protection. During the night, when exposure is minimal, cells undertake DNA repair. To ensure cycle optimization, the genes regulating the circadian rhythm need protection from external damage caused by UV exposure. Day protection Control Chronogen (1%) (before UV irradiation) UVB (60 mJ.cm-2) Cell Culture: Fibroblasts Product Application: Chronogen (1%) once a day Protocol: - Application of Chronogen (1%) for 24 hours - UVB (60 mJ.cm-2) - Evaluation of DNA damage by comet assay 1 hour after UV irradiation The average measure of the tail moment length, representative of the amount of DNA damage, was 78% less when Chronogen was applied to the cells prior to exposure compared to the control. In-vitro results suggest that Chronogen can help boost the natural DNA protection from UV radiation. Night repair Control UVB (60 mJ.cm-2) Chronogen (1%) (after UV irradiation) Cell Culture: Fibroblasts Product Application: Chronogen (1%) once a day Protocol: - UVB (60 mJ.cm-2) - Application of Chronogen (1%) for 24 hours - Evaluation of DNA damage by comet assay The average measure of the tail moment length, representative of the amount of DNA damage, was 81% less when Chronogen was applied to the cells for 24 hours after exposure compared to the control. In-vitro results suggest that Chronogen can help cells better repair DNA from UV damage. In-vivo Skin Protection and Regeneration Two double-blind studies were completed. Each was carried out over 17 days on 12 volunteers (age 24-37) using a cosmetic formulation containing Chronogen (1%) vs. a placebo. A standard quantity of each (2 mg.cm-2) was applied on the thighs twice a day, in the morning and evening. For the first study, the treated areas were exposed to full solar spectrum irradiation (2 MED) on day 15. Twenty four hours later (day 16) an observation was made using confocal microscopy (Vivascope®), with the number of sunburn cells (SBC) counted. The areas treated with the Chronogen formulation had 38% fewer SBCs compared to the areas treated with the placebo, confirming the previously observed in-vitro and ex-vivo results which suggest that Chronogen may help boost the skin’s natural defenses against UV irradiation. In the second study, the stratum corneum renewal was measured using a dihydroxyacetone (DHA) method that stains proteins. Here, DHA was applied on two symmetric zones of the thighs at day 0, and photos and grading were recorded at days 1, 4, 7, 11, 14 and 16. For 11 out of the 12 volunteers, the area treated with the Chronogen formulation was observed to have a faster fading of the DHA staining compared to the placebo treated area, with the best effects noted at day 4 (75% of the volunteers) and day 7 (67% of volunteers). Since skin turnover is directly proportional to the disappearance of the skin color (brown/orange) caused by the application of DHA, the results suggest that Chronogen may help to boost skin turnover and regeneration. Specifications Form: Color: Odor: Solubility: Preservative: Limpid solution Colorless Characteristic Water soluble Phenoxyethanol, Sodium Benzoate INCI Name Water (Aqua), Butylene Glycol, Tetrapeptide-26 Formulation Guidelines – Add in water phase below 40°C; in emulsion add on cool-down post-emulsion – Recommended use levels: 1% (clinically tested at 1%) The above formulation information is for guideline purposes only. Some products may be formulated outside of the parameters of these guidelines. Please contact ISP for more information. WORLD HEADQUARTERS INTERNATIONAL SPECIALTY PRODUCTS 1361 Alps Road, Wayne, New Jersey 07470, USA Tel: +1 973 628-4000 Fax: +1 973 628-3311 www.ispcorp.com [email protected] CUSTOMER SERVICE: Toll Free: 1 (800) 622-4423 Fax: +1 973 628-4001 SAMPLE CENTER: Toll Free: 1 (800) 243-6788 [email protected] REGIONAL SALES OFFICES NORTH AMERICA WAYNE, NEW JERSEY Toll Free: 1 (800) 505-8984 Fax:+1 973-628-4117 [email protected] GLOBAL SKIN RESEARCH CENTRE SOPHIA ANTIPOLIS, FRANCE 655, Route du Pin Montard, BP 212 06904 Sophia Antipolis Cédex, France Tel: +33 (0) 4 92 94 16 06 Fax: +33 (0) 4 92 94 16 00 [email protected] LATIN AMERICA SÃO PAULO, BRAZIL Tel: +55 11 3649-0420 +55 11 3649-0455 +55 11 3649-0468 [email protected] EUROPE COLOGNE, GERMANY Tel: +49 (0) 2236 9649-260 +49 (0) 2236 9649-264 +49 (0) 2236 9649-266 Fax: +49 (0) 2236 9649-295 [email protected] ASIA PACIFIC SOUTHPOINT, SINGAPORE Tel: +656 223-3778 Fax: +656 226-0853 [email protected] ™ Trademark registration applied for ® Registered trademark of the ISP group © International Specialty Products. 2010 Printed in USA. The information contained in this brochure and the various products described are intended for use only by persons having technical skill and at their own discretion and risk after they have performed necessary technical investigations, tests and evaluations of the products and their uses. This material is for informational purposes only and describes the scientific support for the use of Chronogen™ as an ingredient in cosmetic products intended to enhance appearance and other cosmetic benefits. While the information herein is believed to be reliable, we do not guarantee its accuracy and a purchaser must make its own determination of a product’s suitability for purchaser’s use, for the protection of the environment, and for the health and safety of its employees and the purchasers of its products. Neither ISP nor its affiliates shall be responsible for the use of this information, or of any product, method, formulation, or apparatus described in this brochure. Nothing herein waives any of ISP’s or its affiliates’ conditions of sale, and WE MAKE NO WARRANTY, EXPRESS OR IMPLIED, OF MERCHANTABILITY OR FITNESS OF ANY PRODUCT FOR A PARTICULAR USE OR PURPOSE. We also make no warranty against infringement of any patents by reason of purchaser’s use of any information, product, method or apparatus described in this brochure. ispcorp.com IN