Screening of natural molecules of cosmetic interest by combining analytical, bioinformatic and metabolomic tools

Les produits naturels d'origine végétale sont devenus une source majeure de composés bioactifs pour les industries cosmétiques. Cependant, la détermination du principe actif, c'est-à-dire l'identification, l'isolement et le test de chaque molécule, est complexe, longue et fastidieuse compte tenu de la diversité moléculaire des extraits. Dans ce contexte, ces travaux de thèse portent sur la combinaison d’outils analytiques, bioinformatiques et métabolomiques dans le but de développer des approches simples, rapides et efficaces pour cibler et identifier des molécules d’intérêt. Par conséquent, ces travaux ont conduit d’abord à la mise en place de réseaux moléculaires quantitatifs afin d’étudier l’accumulation de métabolites spécialisés dans des cals cellulaires de lin. Ensuite, au développement d’une étude métabolomique par UHPLC-HRMS, associée aux informations de déréplication et de similarités structurales obtenues par les réseaux moléculaires afin de mettre en évidence les variations chimiques par des analyses statistiques, et de caractériser les composés discriminants de deux extraits de plante à différents stades de développement. Enfin, à l’association des réseaux moléculaires et du fractionnement des extraits par chromatographie de partage centrifuge pour la caractérisation phytochimique de deux extraits de plante ayant montré des propriétés d’intérêt pour des applications cosmétiques par des tests chimiques, enzymatiques et cellulaires. Cette combinaison a permis d’obtenir et de tester des fractions simplifiées facilitant ainsi le criblage et l'identification des molécules responsables de l’activité biologique des extraits de plante étudiés.Natural products of plant origin have emerged a major source of bioactive compounds for the cosmetic industry. However, the determination of the active ingredient, i.e. identification, isolation and testing of each molecule, is complex, long and tedious taking into account molecular diversity of the extracts. In this context, this PhD work focuses on the combination of analytical, bioinformatic and metabolomic tools in order to develop simple, rapid and efficient approaches to target and identify molecules of interest. Consequently, this work initially conduced to the implementation of quantitative molecular networks in order to study the accumulation of specialized metabolites in flax cell callus. Then, to develop a metabolomic study by UHPLC-HRMS, associated with the information of dereplication and structural similarities obtained by molecular networks in order to highlight chemical variations by statistical analyses, and to characterize the discriminating compounds of two plants extracts at different stages of development. Finally, to establish a combination of molecular networks and fractionation by centrifugal partition chromatography for the phytochemical characterization of two plant extracts having shown biological properties interesting for cosmetic applications by chemical, enzymatic and cellular assays. This combination made it possible to obtain and test simplified fractions thus facilitating the screening and identification of the molecules responsible for the biological activity of studied plant extracts

Criblage de molécules naturelles d’intérêt cosmétique par combinaison d’outils analytiques, bioinformatiques et métabolomiques

Natural products of plant origin have emerged a major source of bioactive compounds for the cosmetic industry. However, the determination of the active ingredient, i.e. identification, isolation and testing of each molecule, is complex, long and tedious taking into account molecular diversity of the extracts. In this context, this PhD work focuses on the combination of analytical, bioinformatic and metabolomic tools in order to develop simple, rapid and efficient approaches to target and identify molecules of interest. Consequently, this work initially conduced to the implementation of quantitative molecular networks in order to study the accumulation of specialized metabolites in flax cell callus. Then, to develop a metabolomic study by UHPLC-HRMS, associated with the information of dereplication and structural similarities obtained by molecular networks in order to highlight chemical variations by statistical analyses, and to characterize the discriminating compounds of two plants extracts at different stages of development. Finally, to establish a combination of molecular networks and fractionation by centrifugal partition chromatography for the phytochemical characterization of two plant extracts having shown biological properties interesting for cosmetic applications by chemical, enzymatic and cellular assays. This combination made it possible to obtain and test simplified fractions thus facilitating the screening and identification of the molecules responsible for the biological activity of studied plant extracts.Les produits naturels d'origine végétale sont devenus une source majeure de composés bioactifs pour les industries cosmétiques. Cependant, la détermination du principe actif, c'est-à-dire l'identification, l'isolement et le test de chaque molécule, est complexe, longue et fastidieuse compte tenu de la diversité moléculaire des extraits. Dans ce contexte, ces travaux de thèse portent sur la combinaison d’outils analytiques, bioinformatiques et métabolomiques dans le but de développer des approches simples, rapides et efficaces pour cibler et identifier des molécules d’intérêt. Par conséquent, ces travaux ont conduit d’abord à la mise en place de réseaux moléculaires quantitatifs afin d’étudier l’accumulation de métabolites spécialisés dans des cals cellulaires de lin. Ensuite, au développement d’une étude métabolomique par UHPLC-HRMS, associée aux informations de déréplication et de similarités structurales obtenues par les réseaux moléculaires afin de mettre en évidence les variations chimiques par des analyses statistiques, et de caractériser les composés discriminants de deux extraits de plante à différents stades de développement. Enfin, à l’association des réseaux moléculaires et du fractionnement des extraits par chromatographie de partage centrifuge pour la caractérisation phytochimique de deux extraits de plante ayant montré des propriétés d’intérêt pour des applications cosmétiques par des tests chimiques, enzymatiques et cellulaires. Cette combinaison a permis d’obtenir et de tester des fractions simplifiées facilitant ainsi le criblage et l'identification des molécules responsables de l’activité biologique des extraits de plante étudiés

Untargeted metabolomic and molecular network approaches to reveal tomato root secondary metabolites

International audienceIntroduction The tomato plant, Solanum lycopersicum L. (Solanaceae), is one of the most widely consumed vegetables in the world and plays an important role in human diet. Tomato cultivars are hosts for diverse types of pests, implying diverse chemical defence strategies. Glycoalkaloids are the main specialised metabolites produced by tomato leaves and fruits to protect against pests. However, the roots have received little attention, leading to limited knowledge about their phytochemical content.Objective The main goal of the current study was the development of an untargeted ultra-high-performance liquid chromatography high-resolution mass spectrometry (UHPLC-HRMS) based metabolomic approach to study phytochemical variations in tomato roots at two different development stages (i.e. 34th and 62nd day after sowing).Methods UHPLC-HRMS was used to establish the fingerprint of 24 batches of tomato roots. Statistical analyses were performed to highlight the compounds that discriminated between young and mature tomato roots. A dereplication strategy using molecular networking and HRMS/MS data was set up to identify the metabolites regulated during early root development.Key findings The main biomarkers were guanidine and adenosine derivatives associated with tryptophan. Secondary metabolites such as glycoalkaloids and steroidal alkaloids were also characterised. Most of the metabolites were up-regulated in young tomato roots (34 days old) while tryptophan was up-regulated in the older roots (62 days old).Conclusion The metabolic changes observed in this work contribute to a deeper understanding of early-stage root development and may help our understanding of the complex processes involved in the tomato root defence arsenal

UPLC-HRMS Analysis Revealed the Differential Accumulation of Antioxidant and Anti-Aging Lignans and Neolignans in In Vitro Cultures of Linum usitatissimum L

International audienceOver the last few decades, methods relating to plant tissue culture have become prevalent within the cosmetic industry. Forecasts predict the cosmetic industry to grow to an annual turnover of around a few hundred billion US dollars. Here we focused onLinum usitatissimumL., a plant that is well-known for its potent cosmetic properties. Following the a) establishment of cell cultures from three distinct initial explant origins (root, hypocotyl, and cotyledon) and b) selection of optimal hormonal concentrations, twoin vitrosystems (callusvscell suspensions) were subjected to different light conditions. Phytochemical analysis by UPLC-HRMS not only confirmed high (neo)lignan accumulation capacity of this species with high concentrations of seven newly described (neo)lignans. Evaluation over 30 days revealed strong variations between the two differentin vitrosystems cultivated under light or dark, in terms of their growth kinetics and phytochemical composition. Additionally, antioxidant (i.e.four differentin vitroassays based on hydrogen-atom transfer or electron transfer mechanism) and anti-aging (i.e.fourin vitroinhibition potential of the skin remodeling enzymes: elastase, hyaluronidase, collagenase and tyrosinase) properties were evaluated for the two differentin vitrosystems cultivated under light or dark. A prominent hydrogen-atom transfer antioxidant mechanism was illustrated by the DPPH and ABTS assays. Potent tyrosinase and elastase inhibitory activities were also observed, which was strongly influenced by thein vitrosystem and light conditions. Statistical treatments of the data showed relationship of some (neo)lignans with these biological activities. These results confirmed the accumulation of flax (neo)lignans in differentin vitrosystems that were subjected to distinct light conditions. Furthermore, we showed the importance of optimizing these parameters for specific applications within the cosmetic industry