Cécile S. Le Duff

Stability of meropenem and doripenem solutions for administration by continuous infusion

the same pI—despite the negative diffusion test for ESBL detection—we carried out a search for bla SHV using PCR (see Table S2); this, however, was negative for both strains. Another possible explanation for the aztreonam resistance observed in our strains might be overproduction of their chromosomal class A OXY b-lactamase. 5 The promoter region of the OXY b-lactamase was amplified by PCR (see Table S2), and the sequences revealed a mutation in the 210 consensus region of the promoter in both strains; this consisted of the transition (G!A) of the fifth base, described as the most frequent among in vitro mutants and clinical isolates of aztreonam-resistant K. oxytoca. 5 The bla OXY gene was also amplified by PCR (see Table S2), and the sequence analysis indicated that both strains carried a bla OXY-2-type very similar to bla OXY-2-8 (GenBank accession no. AY055205) with only two substitutions, a serine to glycine at position 23 and an aspartic acid to alanine at position 38, considering position 1 as the starting methionine.

Self-Assembling Doxorubicin−Tocopherol Succinate Prodrug as a New Drug Delivery System: Synthesis, Characterization, and in Vitro and in Vivo Anticancer Activity

Self-assembled prodrugs forming nanoaggregates are a promising approach to enhance the antitumor efficacy and to reduce the toxicity of anticancer drugs. To achieve this goal, doxorubicin was chemically conjugated to d-α-tocopherol succinate through an amide bond to form N-doxorubicin-α-d-tocopherol succinate (N-DOX-TOS). The prodrug self-assembled in water into 250 nm nanostructures when stabilized with d-α-tocopherol poly(ethylene glycol) 2000 succinate. Cryo-TEM analysis revealed the formation of nanoparticles with a highly ordered lamellar inner structure. NMR spectra of the N-DOX-TOS nanoparticles indicated that N-DOX-TOS is located in the core of the nanoparticles while PEG chains and part of the tocopherol are in the corona. High drug loading (34% w/w) and low in vitro drug release were achieved. In vitro biological assessment showed significant anticancer activity and temperature-dependent cellular uptake of N-DOX-TOS nanoparticles. In vivo, these nanoparticles showed a greater antitumor efficacy than free DOX. N-DOX-TOS nanoparticles might have the potential to improve DOX-based chemotherapy.

Induction of Highly Curved Structures in Relation to Membrane Permeabilization and Budding by the Triterpenoid Saponins, α- and δ-Hederin

Background: The triterpenoid monodesmosidic saponins, α- and δ-hederin, induced membrane permeabilization. Results: The membranous cholesterol and the sugars branched on the aglycone, hederagenin, are critical for membrane permeabilization, budding, and the change in lipid phase. Conclusion: Permeabilization and budding are dependent on the interaction of saponin with cholesterol and the molecular shape of the saponin. Significance: Induction of curvature by saponins is responsible for permeabilization and budding. The interactions of triterpenoid monodesmosidic saponins, α-hederin and δ-hederin, with lipid membranes are involved in their permeabilizing effect. Unfortunately, the interactions of these saponins with lipid membranes are largely unknown, as are the roles of cholesterol or the branched sugar moieties (two for α-hederin and one for δ-hederin) on the aglycone backbone, hederagenin. The differences in sugar moieties are responsible for differences in the molecular shape of the saponins and the effects on membrane curvature that should be the most positive for α-hederin in a transbilayer direction. In large unilamellar vesicles and monocyte cells, we showed that membrane permeabilization was dependent on the presence of membrane cholesterol and saponin sugar chains, being largest for α-hederin and smallest for hederagenin. In the presence of cholesterol, α-hederin induced the formation of nonbilayer phases with a higher rate of Brownian tumbling or lateral diffusion. A reduction of Laurdans generalized polarization in relation to change in order of the polar heads of phospholipids was observed. Using giant unilamellar vesicles, we visualized the formation of wrinkled borders, the decrease in liposome size, budding, and the formation of macroscopic pores. All these processes are highly dependent on the sugars linked to the aglycone, with α-hederin showing a greater ability to induce pore formation and δ-hederin being more efficient in inducing budding. Hederagenin induced intravesicular budding but no pore formation. Based on these results, a curvature-driven permeabilization mechanism dependent on the interaction between saponin and sterols and on the molecular shape of the saponin and its ability to induce local spontaneous curvature is proposed.

Effects of Thickness and Grafting Density on the Activity of Polymer-Brush-Immobilized Tris(triazolyl) Copper(I) Catalysts

We report herein the application of polymer brushes for the immobilization of tris[(1,2,3‐triazolyl)methyl]amine CuCl catalysts. Well‐defined catalytic brushes were prepared through grafting‐from and postpolymerization modification approaches on Si surfaces and characterized by X‐ray reflectivity, X‐ray photoelectron spectroscopy, and inductively coupled plasma mass spectrometry. Hairy catalysts of varying thickness and grafting density were investigated in a model CuI‐catalyzed azide–alkyne cycloaddition reaction and showed remarkable activity at loadings as low as 0.02 mol % as a result of the unique catalytic site density and nuclearity found in the brush. We demonstrate that thickness and grafting density parameters can be adjusted to maximize catalytic activity along the brush thickness.

The Blood Flow Shutdown Induced by Combretastatin A4 Impairs Gemcitabine Delivery in a Mouse Hepatocarcinoma

In recent clinical studies, vascular disrupting agents (VDAs) are mainly used in combination with chemotherapy. However, an often overlooked concern in treatment combination is the VDA-induced impairment of chemotherapy distribution in the tumor. The work presented here investigated the impact of blood flow shutdown induced by Combretastatin A4 (CA4) on gemcitabine uptake into mouse hepatocarcinoma. At 2 h after CA4 treatment, using DCE-MRI, a significant decrease in the perfusion-relevant parameters Ktrans and Vp were observed in treated group compared with the control group. The blood flow shutdown was indeed confirmed by a histology study. In a third experiment, the total gemcitabine uptake was found to be significantly lower in treated tumors, as assessed in a separate experiment using ex vivo fluorine nuclear magnetic resonance spectroscopy. The amount of active metabolite gemcitabine triphosphate was also lower in treated tumors. In conclusion, the blood flow shutdown induced by VDAs can impact negatively on the delivery of small cytotoxic agents in tumors. The present study outlines the importance of monitoring the tumor vascular function when designing drug combinations.