Final Report Summary - NATPROTEC (Integrated technologies for the discovery and development of cosmeceutical agents from plant biodiversity) NATPROTEC project aimed to discover and carry to the stage of development innovative products in the area of cosmeceuticals originating from European natural resources using emerging and environmentally friendly technologies. The project launched on the 1rst of February 2012 and had duration of 4 years. Specifically, NATPROTEC’s scientific concept involved the discovery of novel natural products originating from the Mediterranean and Alpine biodiversity. In parallel, the competiveness of European industries was expanded in the area of natural products chemistry and biology with cosmeceutical application which is considered as an emerging field of research and development. The key objectives were: • Utilization of existing structurally diverse libraries of natural products to serve as a pre-selection platform for the focused discovery of scaffolds acting as cosmeceutical agents using high throughput in silico and in vitro methods• Exploitation of the Alpine and Mediterranean biodiversity for the discovery of novel candidates to be developed as possible cosmeceuticals• Development, optimization and application of emerging “green” extraction methods (SFE-CO2, SWE, ART, CPE) and incorporation of state-of–the-art miniaturized approaches and advanced analytical techniques (microfractionation, cap-NMR, LC-SPE-NMR, LC-HR-MSn) for the bioactivity-oriented isolation and structure elucidation of the active constituents• Application of a comprehensive panel of bioassays and integration of metabolomic approaches for the evaluation of cosmeceutical properties of the active agents• Development of innovative lead products for cosmeceutical applications and utilization of metabolic profiling techniques (HPTLC-, UV-, MS-, NMR-based) for the optimization and quality assessment thereof• Exploitation of emerging techniques, novel approaches and advanced methodologies produced in the academic environment for the benefit of the companies involved and establishment of dynamic synergies between Industry and Academia through exploitation of complimentary expertise and mutual understanding in different environments and skills • Enhancement of the competitiveness of the European industries in the area of cosmeceuticals and broadening the potentialities for the development of optimized products in favor of the consumers and the society These objectives were implemented through an extended and balanced scheme of researcher’s exchanges and recruitments, in both directions and via a mutual scientific project developed on the needs and interests of both Industry and Academia sectors. A high-calibre network was established including four European academic entities and four industrial partners. National and Kapodistrian University of Athens (Greece), University of Geneva (Switzerland), University of Innsbruck (Austria) and University of Reims Champagne-Ardenne (France) are universities with documented high quality research activity in the field of natural products. Rousslet Centifugation SA (France) is an internationally leading industry in Centrifugal Partition Chromatography (CPC) technology, KORRES SA Natural Products (Greece) is a rapidly growing company recognized for its expertise in natural cosmetics, Bionorica Reserach GmbH (Austria) specializes in innovative analytical approaches and CAMAG (Switzerland) is renowned worldwide for its unique expertise in High Performance Thin Layer Chromatography (HPTLC). Mediterranean and Alpine biodiversities were rationally exploited based on in silico calculations of existing compounds libraries, in vitro evaluation as well as literature and empirical knowledge in the context of cosmetic applications. Thus, a list with the most interesting plants to be studied was formed. The plants were collected and extracted and a broad spectrum of bioassays was employed. For the evaluation of skin-protective and anti-hyperpigmentation effects of the extracts the following assays were performed: against histone deacetylase (HDAC) activity, antioxidant activity, quinone reductase (QR) induction activity and tyrosinase inhibition activity. The results were combined, filtered and 26 extracts with significant activity were selected for further investigation. Extracts with promising anti-hyperpigmentation activity were further checked for their anti-ageing properties (inhibition of elastase, hyaluronidase and collagenase activity). In parallel, new bioevaluation methodologies were developed. Bioautography assay for detection of tyrosinase inhibitors was optimized and validated on HPTLC. Additionally, a strategy was implemented on UHPLC-ToF-MS in combination with multivariate analysis, to study metabolic changes in quinone reductase (NQO1) plant extract treated HeLa cells. The 26 selected plants-extracts were forwarded to microfractionation and phytochemical study employing state-of-the-art analytical techniques. Biological activity of fractions was correlated to specific compounds that were further proceeded to bioevaluation. The efficient, accelerated and advantageous isolation and identification of natural constituents was performed mainly with centrifugal partition chromatography. Among the plants studied, Crocus sativus, Leucanthemum vulgare, Olea europaea and Pistacia lentiscus were the four plants that distinguished and were selected for development. The optimal raw plant material leading to the most promising extracts in terms of biological activity and metabolic profile was selected. Conditions of extraction were optimized, focusing on “green” technologies (SFE-CO2, resins, CPE) and the extracts were forwarded to pilot scale. Formulation technology in combination with market interest depicted the extract produced from Crocus sativus stigmas as the most appropriate for incorporation into the final cosmeceutical product. Within NATPROTEC, core scientific knowledge and lead compounds that could be further exploited were developed. Apart from the cosmeceutical product, new environmentally friendly extraction techniques, new analytical methods for the characterisation of plant extracts and isolation and identification of natural products, new validated analytical protocols for the quality assessment of the final products and new bioautographic assays for the high throughput biological evaluation of natural products were developed. In parallel, valuable synergies were created through implemented secondments and recruitments. NATPROTEC project strengthened the interaction between Industry and Academia, enhanced their cooperation and the interchange of knowledge for dual benefit. Please find further information on http://www.natprotec.eu/or please contact Prof. Leandros Skaltsounisskaltsounis@pharm.uoa.grTel. +30 210 727 45 98Fax. +30 210 727 45 94
