Laboratory of Design and Application of Bioactive Molecules

The growth of hyperbranched polymers by self-condensing vinyl polymerization under RAFT conditions from the surface of upconversion nanoparticles is hindered by steric hinderance, but also increased termination and transfer reactions.

o-Nitrobenzyl (o-NB) derivatives are the most widely applied photoremovable groups for the study of dynamic biological processes. By introducing different substituents to the benzylic position we were able to generate a fluorescence signal upon irradiation. This signal originates from the formation of a nitrosoketone by-product able to achieve a keto-enol tautomerism leading to pi-conjugated α-hydroxystilbene derivatives. These o-NB caging groups can be used to directly monitor the uncaging event by the release of a detectable fluorescent side-product.

We obtained osmotic pressure data of lysozyme solutions, describing their physical states over a wide concentration range, using osmotic stress for pressures between 0.05 bar and about 40 bar and volume fractions between 0.01 and 0.61. The osmotic pressure vs. volume fraction data consist of a dilute, gas-phase regime, a transition regime with a high-compressibility plateau, and a concentrated regime where the system is nearly incompressible. The first two regimes are shifted towards a higher protein volume fraction upon decreasing the strength or the range of electrostatic interactions. We describe this shift and the overall shape of the experimental data in these two regimes through a model accounting for a steric repulsion, a short-range van der Waals attraction and a screened electrostatic repulsion. The transition is caused by crystallization, as shown by small-angle X-ray scattering. We verified that our data points correspond to thermodynamic equilibria, and thus that they consist of the reference experimental counterpart of a thermodynamic equation of state.

In this study, we propose a new solution for the nanoencapsulation of hydrophilic anticancer drug, doxorubicin hydrochloride (DOX). The drug molecules are solubilized in the core of aqueous nanoreservoirs, so-called aqueous core nanocapsules (ACN) recently developed by our team, and dispersed in aqueous bulk media. Since it is well acknowledged that the nanoencapsulation of DOX has many advantages, like reducing the sides effects (e.g. cardiac toxicity), we propose through the present study a novel formulation solution for this purpose. After focusing on the formulation process for optimizing the drug encapsulation yield, the DOX-release profiles were followed up and analyzed. Different physicochemical and in vitro characterization were performed, and complement activation experiments. ACN were shown efficient to encapsulate DOX reaching yields as high as 80%, followed by a sustained release governed by a diffusion-controlled mechanism. The loaded nanocarriers showed low levels of complement activation, compatible with stealth properties. To summarize, this study brings out a new tool for the nanoencapsulation of hydrophilic anticancers and could open new doors for the administration of this particular class of drugs.

Size-tunable polymeric nanoparticles have been successfully produced by a microfluidic-assisted nanoprecipitation process. A multilamination micromixer has been chosen to fabricate continuously nanoparticles of methacrylic polymers. Various operating conditions, such as the polymer concentration, the amount of non-solvent and the characteristics of the raw polymer (molecular weight and architecture: linear vs. branched) have been investigated. Their influences on the final particle size, ranging from 76 to 217 nm, have been correlated to the mechanisms leading to the formation of nanoparticles. In this type of microfluidic device, mixing mainly operates by diffusion mass transfer, helped by hydrodynamic focusing. The effect of micromixing on the size of particles has also been shown experimentally and supported by a computational fluid dynamics (CFD) study. A mixing criterion has been defined and numerically calculated to corroborate the effect of the flow rate of polymer solution on the particles size. An increase in the polymer solution flow rate increases the value of this mixing criterion, resulting in smaller nanoparticles.

Lipophilic and axially substituted tri-n-hexylsiloxy aluminum phthalocyanine and cholesteryloxy diphenylsiloxy aluminum phthalocyanine were synthesized and assayed in PDT against M6 melanocytes. In the conditions used (λ > 480 nm , 10 mW cm-2, egg-yolk lecithin or cremophor EL formulation) they both exhibited a higher photodynamic effect than chloroaluminum phthalocyanine. They displayed 2% to 3.5% cell viability at 10-5M dose for 20 min irradiation. Hexadecafluoro zinc phthalocyanine was synthesized to increase the lipophilicity of zinc phthalocyanine, hexadecachloro zinc phthalocyanine was also included because it would theoretically enhance the phototoxicity. In all the delivery systems used, their photodynamic effect against M6 melanocytes was lower in comparison with zinc phthalocyanine and axially substituted aluminum phthalocyanines. A 2 h irradiation treatment with 3 × 10-6 M hexadecafluoro zinc phthalocyanine and 10-5 M hexadecachloro zinc phthalocyanine led to 60% and 15% cell viability respectively. In all cases, the cell killing effect was light-and dose-dependent and was higher in cremophor EL micelles than in the egg-yolk lecithin formulation.

Helichrysum stoechas L. plants were hydrodistilled and the oil, which was obtained in 0.04% yield, was analyzed by GC and GC/MS. The major components were α-pinene (63%), limonene (14%), β-caryophyllene (2.8%) and α-humulene (2%). Other separated components (≃90) accounted for less than 10% of the oil.