Etude de la synthèse de polyamides par catalyse enzymatique et de la résolution cinétique dynamique de diamines : une approche de la synthèse de polyamides chiraux par catalyse tandem itérative

La catalyse tandem itérative permet la synthèse de polyesters chiraux à partir de monomères racémiques, en associant one-pot une polymérisation enzymatique énantiosélective et une réaction de racémisation. L ensemble de ce travail constitue une étude préliminaire à l extension de cette méthodologie aux polyamides. La capacité de l enzyme CAL-B à catalyser la synthèse de polyamides solubles de masses moléculaires moyennes élevées a été démontrée. Les polymères isolés ont pu être pleinement caractérisés. Cette méthodologie a pu être étendue à la synthèse de polyamides chiraux à partir de monomères optiquement actifs. La réaction de résolution cinétique dynamique associant résolution enzymatique et racémisation par les radicaux sulfanyle a ensuite été étendue au contrôle de deux centres asymétriques, via l utilisation de diamines C2-symétriques. Cette méthodologie a pu être utilisée avec la même efficacité à 40 et à 80C.AIX-MARSEILLE3-BU Sc.St Jérô (130552102) / SudocSudocFranceF

Efficient Ocular Delivery of VCP siRNA via Reverse Magnetofection in RHO P23H Rodent Retina Explants

The use of synthetic RNA for research purposes as well as RNA-based therapy and vaccination has gained increasing importance. Given the anatomical seclusion of the eye, small interfering RNA (siRNA)-induced gene silencing bears great potential for targeted reduction of pathological gene expression that may allow rational treatment of chronic eye diseases in the future. However, there is yet an unmet need for techniques providing safe and efficient siRNA delivery to the retina. We used magnetic nanoparticles (MNPs) and magnetic force (Reverse Magnetofection) to deliver siRNA/MNP complexes into retinal explant tissue, targeting valosin-containing protein (VCP) previously established as a potential therapeutic target for autosomal dominant retinitis pigmentosa (adRP). Safe and efficient delivery of VCP siRNA was achieved into all retinal cell layers of retinal explants from the RHO P23H rat, a rodent model for adRP. No toxicity or microglial activation was observed. VCP silencing led to a significant decrease of retinal degeneration. Reverse Magnetofection thus offers an effective method to deliver siRNA into retinal tissue. Used in combination with retinal organotypic explants, it can provide an efficient and reliable preclinical test platform of RNA-based therapy approaches for ocular diseases

Self-Amplifying Replicon RNA Delivery to Dendritic Cells by Cationic Lipids

Advances in RNA technology during the past two decades have led to the construction of replication-competent RNA, termed replicons, RepRNA, or self-amplifying mRNA, with high potential for vaccine applications. Cytosolic delivery is essential for their translation and self-replication, without infectious progeny generation, providing high levels of antigen expression for inducing humoral and cellular immunity. Synthetic nanoparticle-based delivery vehicles can both protect the RNA molecules and facilitate targeting of dendritic cells—critical for immune defense development. Several cationic lipids were assessed, with RepRNA generated from classical swine fever virus encoding nucleoprotein genes of influenza A virus. The non-cytopathogenic nature of the RNA allowed targeting to dendritic cells without destroying the cells—important for prolonged antigen production and presentation. Certain lipids were more effective at delivery and at promoting translation of RepRNA than others. Selection of particular lipids provided delivery to dendritic cells that resulted in translation, demonstrating that delivery efficiency could not guarantee translation. The observed translation in vitro was reproduced in vivo by inducing immune responses against the encoded influenza virus antigens. Cationic lipid-mediated delivery shows potential for promoting RepRNA vaccine delivery to dendritic cells, particularly when combined with additional delivery elements. Keywords: replicon, RepRNA, self-amplifying, cationic lipids, dendritic cells, nanoparticle delivery, influenza vaccines, humoral immunity, cellular immunit

Synthesis and properties of a series of beta-cyclodextrin/nitrone spin traps for improved superoxide detection

International audienceThree new DEPMPO-based spin traps have been designed and synthesized for improved superoxide detection, each carrying a cyclodextrin (CD) moiety but with a different alkyl chain on the phosphorus atom or with a long spacer arm. EPR spectroscopy allowed us to estimate the half-life of the superoxide spin adducts which is close to the value previously reported for CD-DEPMPO (t(1/2) approximate to 50-55 min under the conditions investigated). The spectra are typical of superoxide adducts (almost no features of the HO center dot adduct that usually forms with time for other nitrone spin traps such as DMPO) and we show that at 250 mu M, the new spin trap enables the reliable detection of superoxide by 1 scan at the position opposite to the corresponding spin trap without the CD moiety. The resistance of the spin adducts to a reduction process has been evaluated, and the superoxide spin adducts are sensitive to ascorbate and glutathione (GSH), but not to glutathione peroxidase/GSH, reflecting the exposed nature of the nitroxide moiety to the bulk solvent. To understand these results, 2D-ROESY NMR studies and molecular dynamics pointed to a shallow or surface self-inclusion of the nitrone spin traps and of nitroxide spin adducts presumably due to the high flexibility of the permethyl-beta-CD rim