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46 results on '"Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)"'

1. Etching of semiconductors and metals by the photonic jet with shaped optical fiber tips

Author

Pierre Pfeiffer, Robin Pierron, Joël Fontaine, Sylvain Lecler, Frédéric Mermet, Julien Zelgowski, Laboratoire des sciences de l'ingénieur, de l'informatique et de l'imagerie (ICube), Institut National des Sciences Appliquées - Strasbourg (INSA Strasbourg), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-École Nationale du Génie de l'Eau et de l'Environnement de Strasbourg (ENGEES)-Réseau nanophotonique et optique, Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Matériaux et nanosciences d'Alsace (FMNGE), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), IREPA Laser [Illkirch, France] (Pôle API), Institut Carnot MICA, Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), École Nationale du Génie de l'Eau et de l'Environnement de Strasbourg (ENGEES)-Université de Strasbourg (UNISTRA)-Institut National des Sciences Appliquées - Strasbourg (INSA Strasbourg), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National de Recherche en Informatique et en Automatique (Inria)-Les Hôpitaux Universitaires de Strasbourg (HUS)-Centre National de la Recherche Scientifique (CNRS)-Matériaux et Nanosciences Grand-Est (MNGE), Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Réseau nanophotonique et optique, Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), and Pierron, Robin

Subjects
[PHYS.PHYS.PHYS-OPTICS] Physics [physics]/Physics [physics]/Optics [physics.optics], Optical fiber, Materials science, Silica fiber, Silicon, Laser etching, Optical ber, Energy balance, FOS: Physical sciences, General Physics and Astronomy, chemistry.chemical_element, Applied Physics (physics.app-ph), 02 engineering and technology, Sciences de l'ingénieur [physics]/Autre, 01 natural sciences, law.invention, 010309 optics, Optics, law, 0103 physical sciences, Saturation (magnetic), Photonic jet, [PHYS.PHYS.PHYS-OPTICS]Physics [physics]/Physics [physics]/Optics [physics.optics], business.industry, Physics - Applied Physics, Surfaces and Interfaces, General Chemistry, 021001 nanoscience & nanotechnology, Condensed Matter Physics, Surfaces, Coatings and Films, Micromachining, Semiconductor, chemistry, Metals, Photonics, 0210 nano-technology, business, Titanium
Abstract

International audience; The etching of semiconductors and metals by a photonic jet (PJ) generated with a shaped optical ber tip is studied. Etched marks with a diameter of 1 micron have been realized on silicon, stainless steel and titanium with a 35 kHz pulsed laser, emitting 100 ns pulses at 1064 nm. The selection criteria of the ber and its tip are discussed. We show that a 100/140 silica ber is a good compromise which takes into account the injection, the working distance and the energy coupled in the higherorder modes. The energy balance is performed on the basis of the known ablation threshold of the material. Finally, the dependence between the etching depth and the number of pulses is studied. Saturation is observed probably due to a redeposition of the etched material, showing that a higher pulse energy is required for deeper etchings.

Published
2017

2. Long focal length high repetition rate femtosecond laser glass welding

Author

Kokou D. Dorkenoo, Marion Gstalter, Sylvain Lecler, Jean Luc Rehspringer, A. Bahouka, Grégoire Chabrol, Laboratoire des sciences de l'ingénieur, de l'informatique et de l'imagerie (ICube), Institut National des Sciences Appliquées - Strasbourg (INSA Strasbourg), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-École Nationale du Génie de l'Eau et de l'Environnement de Strasbourg (ENGEES)-Réseau nanophotonique et optique, Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Matériaux et nanosciences d'Alsace (FMNGE), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Institut Carnot MICA, Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Institut de Physique et Chimie des Matériaux de Strasbourg (IPCMS), Université de Strasbourg (UNISTRA)-Matériaux et nanosciences d'Alsace (FMNGE), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS)-Réseau nanophotonique et optique, Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA), ECAM Strasbourg-Europe, École Nationale du Génie de l'Eau et de l'Environnement de Strasbourg (ENGEES)-Université de Strasbourg (UNISTRA)-Institut National des Sciences Appliquées - Strasbourg (INSA Strasbourg), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut National de Recherche en Informatique et en Automatique (Inria)-Les Hôpitaux Universitaires de Strasbourg (HUS)-Centre National de la Recherche Scientifique (CNRS)-Matériaux et Nanosciences Grand-Est (MNGE), Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Réseau nanophotonique et optique, Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Matériaux et Nanosciences Grand-Est (MNGE), and univOAK, Archive ouverte

Subjects
Materials science, [SPI.OPTI] Engineering Sciences [physics]/Optics / Photonic, business.industry, Welding, Laser, 01 natural sciences, Atomic and Molecular Physics, and Optics, Numerical aperture, law.invention, 010309 optics, Lens (optics), Optics, law, Residual stress, 0103 physical sciences, Femtosecond, Rayleigh length, [SPI.OPTI]Engineering Sciences [physics]/Optics / Photonic, Focal length, Electrical and Electronic Engineering, business, Engineering (miscellaneous)
Abstract

A long focal length focusing device is proposed for the process of glass welding by femtosecond laser pulses at high repetition rate and we report on the significant advantages. The study is performed using a 100 mm focusing length F-theta lens. The results are compared to those obtained with a high numerical aperture microscope objective. The long focal length with the associated Rayleigh length method allows a robust high process speed: welding at 1000 mm/s has been achieved, several orders of magnitude larger compared to what was reported until now. Moreover, for one pulse, the same energy on a larger laser spot leads to a lower thermic gradient increase. As a result, with the proposed parameters, the heat accumulation process reduces the residual stress in the welding seams, preventing the formation of fractures inside the seams: mechanical resistance at 30 MPa has been measured.

Published
2019

3. Novel Doxorubicin Derivatives: Synthesis and Cytotoxicity Study in 2D and 3D in Vitro Models

Author

Annie Marc, Thierry F. Vandamme, Pavel S. Chelushkin, Roman Akasov, Daria Zaytseva-Zotova, Elena Markvicheva, Sergey V. Burov, Maria Leko, Natalia L. Klyachko, Isabelle Chevalot, M. Drozdova, Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry (IBCh RAS), Russian Academy of Sciences [Moscow] (RAS), Sechenov First Moscow State Medical University, Institute of Macromolecular Compounds, Laboratoire Réactions et Génie des Procédés (LRGP), Université de Lorraine (UL)-Centre National de la Recherche Scientifique (CNRS), Lomonosov Moscow State University (MSU), Conception et application de molécules bioactives (CAMB), Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), and Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)

Subjects
0301 basic medicine, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, [SDV]Life Sciences [q-bio], Serum albumin, Pharmaceutical Science, Multiple drug resistance, 03 medical and health sciences, medicine, polycyclic compounds, Doxorubicin, General Pharmacology, Toxicology and Pharmaceutics, Cytotoxicity, skin and connective tissue diseases, Microencapsulation, ComputingMilieux_MISCELLANEOUS, biology, Chemistry, lcsh:RM1-950, technology, industry, and agriculture, Aantitumor drug screening assays, Human serum albumin, In vitro, 3. Good health, carbohydrates (lipids), 030104 developmental biology, lcsh:Therapeutics. Pharmacology, Multicellular spheroids, biology.protein, Cancer research, Fetal bovine serum, medicine.drug, Conjugate
Abstract

Purpose: Multidrug resistance (MDR) of tumors to chemotherapeutics often leads to failure of cancer treatment. The aim of the study was to prepare novel MDR-overcoming chemotherapeutics based on doxorubicin (DOX) derivatives and to evaluate their efficacy in 2D and 3D in vitro models. Methods: To overcome MDR, we synthesized five DOX derivatives, and then obtained non-covalent complexes with human serum albumin (HSA). Drug efficacy was evaluated for two tumor cell lines, namely human breast adenocarcinoma MCF-7 cells and DOX resistant MCF-7/ADR cells. Additionally, MCF-7 cells were entrapped in alginate-oligochitosan microcapsules, and generated tumor spheroids were used as a 3D in vitro model to study cytotoxicity of the DOX derivatives. Results: Due to 3D structure, the tumor spheroids were more resistant to chemotherapy compared to monolayer culture. DOX covalently attached to palmitic acid through hydrazone linkage (DOX-N2H-Palm conjugate) was found to be the most promising derivative. Its accumulation levels within MCF-7/ADR cells was 4- and 10-fold higher than those of native DOX when the conjugate was added to cultivation medium without serum and to medium supplemented with 10% fetal bovine serum, respectively. Non-covalent complex of the conjugate with HSA was found to reduce the IC50 value from 32.9 µM (for free DOX-N2H-Palm) to 16.8 µM (for HSA-DOX-N2H-Palm) after 72 h incubation with MCF-7/ADR cells. Conclusion: Palm-N2H-DOX conjugate was found to be the most promising DOX derivative in this research. The formation of non-covalent complex of Palm-N2H-DOX conjugate with HSA allowed improving its anti-proliferative activity against both MCF-7 and MCF-7/ADR cells.

Published
2017

4. Enhancing tumor specific immune responses by transcutaneous vaccination

Author

Vincent Flacher, Sylvie Fournel, Béatrice Heurtault, Soulaima Chamat, Hasnaa Bouharoun-Tayoun, Hanadi Saliba, Benoît Frisch, Conception et application de molécules bioactives (CAMB), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Laboratory of Immunology [Fanar, Lebanon] (Faculty of Public Health - EDST E006), Lebanese University [Beirut] (LU), and Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)

Subjects
0301 basic medicine, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, administration & dosage, Immunology, Tumor specific, Transcutaneous vaccination, [SDV.CAN]Life Sciences [q-bio]/Cancer, Administration, Cutaneous, Cancer Vaccines, methods, Immune tolerance, immunology, 03 medical and health sciences, Immune system, Neoplasms, Drug Discovery, medicine, Stratum corneum, Humans, ComputingMilieux_MISCELLANEOUS, Skin, Pharmacology, therapy, Clinical Trials as Topic, integumentary system, business.industry, Vaccination, Cancer, Dendritic Cells, Sciences du Vivant [q-bio]/Immunologie, medicine.disease, 3. Good health, Treatment Outcome, 030104 developmental biology, medicine.anatomical_structure, Liposomes, [SDV.IMM]Life Sciences [q-bio]/Immunology, Nanoparticles, Molecular Medicine, Cancer vaccine, [SDV.IMM.VAC]Life Sciences [q-bio]/Immunology/Vaccinology, business
Abstract

Our understanding of the involvement of the immune system in cancer control has increased over recent years. However, the development of cancer vaccines intended to reverse tumor-induced immune tolerance remains slow as most current vaccine candidates exhibit limited clinical efficacy. The skin is particularly rich with multiple subsets of dendritic cells (DCs) that are involved to varying degrees in the induction of robust immune responses. Transcutaneous administration of cancer vaccines may therefore harness the immune potential of these DCs, however, this approach is hampered by the impermeability of the stratum corneum. Innovative vaccine formulations including various nanoparticles, such as liposomes, are therefore needed to properly deliver cancer vaccine components to skin DCs. Areas covered: The recent insights into skin DC subsets and their functional specialization, the potential of nanoparticle-based vaccines in transcutaneous cancer vaccination and, finally, the most relevant clinical trial advances in liposomal and in cutaneous cancer vaccines will be discussed. Expert commentary: To define the optimal conditions for mounting protective skin DC-induced anti-tumor immune responses, investigation of the cellular and molecular interplay that controls tumor progression should be pursued in parallel with clinical development. The resulting knowledge will then be translated into improved cancer vaccines that better target the most appropriate immune players. journal article review research support, non-u.s. gov't 2017 11 imported

Published
2017

5. A new {Fe4Co4} soluble switchable nanomagnet encapsulating Cs+: enhancing the stability and redox flexibility and tuning the photomagnetic effect

Author

J. von Bardeleben, Lise-Marie Chamoreau, Yanling Li, M. Tricoire, D. Garnier, Juan-Ramón Jiménez, Yves Journaux, Rodrigue Lescouëzec, Institut Parisien de Chimie Moléculaire (IPCM), Université Pierre et Marie Curie - Paris 6 (UPMC)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Conception et application de molécules bioactives (CAMB), Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Couches nanométriques : formation, interfaces, défauts (INSP-E5), Institut des Nanosciences de Paris (INSP), Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS), and Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Prussian blue, 010405 organic chemistry, Metal ions in aqueous solution, Inorganic chemistry, Photomagnetic effect, Nuclear magnetic resonance spectroscopy, 010402 general chemistry, Photochemistry, 01 natural sciences, Nanomagnet, 0104 chemical sciences, Ion, law.invention, Inorganic Chemistry, chemistry.chemical_compound, chemistry, law, Single-molecule magnet, [CHIM.OTHE]Chemical Sciences/Other, Electron paramagnetic resonance
Abstract

International audience; We report a new cyanide-bridged Cs⊂{Fe4Co4} box, a soluble model of photomagnetic Prussian blue analogues (PBAs). The Cs+ ion has a high affinity for the box and can replace the K+ ion in the preformed K-cube. This exchange is kinetically impeded at room temperature but is accelerated by heating and using the 18-crown-6 ether. The inserted Cs+ ion confers a high robustness to the cube, which withstands boiling, as shown by variable-temperature NMR studies. The stability of this model complex in solution allows the probing of the electronic interaction between the alkali ion and the cyanide cage by using various techniques. These interactions are known to play a role in the photomagnetic behaviour of PBAs. Firstly, the 133Cs NMR spectroscopy proves that there is an electronic communication between the encapsulated alkali ion and the cyanide cage. The measured up-field signal, observed at ca. −200 ppm at 300 K, reveals that a certain amount of spin density is transferred through the bonds from the paramagnetic Co(II) ion to the encapsulated cation. Secondly, cyclovoltammetric studies show that the nature of the inserted ions affects the redox properties of the cage and influences the electronic communication between the metal ions. However, the differences in the electrochemical properties of the K-cube and the Cs-cube remain moderate. As the switching properties are influenced by the redox potential of the Fe and Co centers, similar photomagnetic behaviour is observed, with both of them being highly photomagnetic. This result contrasts strikingly with previous studies on the 3D polymeric PBAs, where the PBAs with a high amount of Cs+ show poor photomagnetic behaviour. In that case, cooperative behaviour likely influences the switching properties. Finally, EPR spectroscopy shows that the K-cube is more anisotropic than the Cs-cube. This difference is reflected in the changes occurring in the slow magnetic relaxation (single molecule magnet behaviour) observed in the two cubes.

Published
2017

6. Self-aggregating 1.8 kDa polyethylenimines with dissolution switch at endosomal acidic pH are delivery carriers for plasmid DNA, mRNA, siRNA and exon-skipping oligonucleotides

Author

Nassera Tounsi, Antoine Kichler, Guy Zuber, Manuela Chiper, Ryszard Kole, Biotechnologie et signalisation cellulaire (BSC), Université de Strasbourg (UNISTRA)-Institut de recherche de l'Ecole de biotechnologie de Strasbourg (IREBS)-Centre National de la Recherche Scientifique (CNRS), Laboratoire d'Innovation Thérapeutique (LIT), Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Conception et application de molécules bioactives (CAMB), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), and Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)

Subjects
mdx mouse, Aucun, Phosphorothioate Oligonucleotides, Pharmaceutical Science, macromolecular substances, 02 engineering and technology, Biology, 010402 general chemistry, 01 natural sciences, chemistry.chemical_compound, [SDV.SP.MED]Life Sciences [q-bio]/Pharmaceutical sciences/Medication, In vivo, Animals, Humans, Polyethyleneimine, RNA, Messenger, RNA, Small Interfering, [SDV.IB.BIO]Life Sciences [q-bio]/Bioengineering/Biomaterials, ComputingMilieux_MISCELLANEOUS, Polyethylenimine, Oligonucleotide, Gene Transfer Techniques, technology, industry, and agriculture, Chemical modification, DNA, Exons, Hydrogen-Ion Concentration, 021001 nanoscience & nanotechnology, Molecular biology, Exon skipping, 0104 chemical sciences, chemistry, Biochemistry, Mice, Inbred mdx, Nucleic acid, [SDV.IB]Life Sciences [q-bio]/Bioengineering, 0210 nano-technology, HeLa Cells, Plasmids
Abstract

Efficiency of polyethylenimine (PEI) for nucleic acid delivery is affected by the size of the carrier and length of the nucleic acids. For instance, PEIs with molecular weights between 10-30kDa provide optimal DNA delivery activity whereas PEIs with molecular weights below 1.8kDa are ineffective. The activity of PEI is also severely diminished by substitution of DNA for shorter nucleic acids such as mRNA or siRNA. Here, through chemical modification of the primary amines to aromatic domains we achieved nucleic acid delivery by the 1.8kDa polyethylenimine (PEI) particles. This modification did not affect the PEI buffering abilities but enhanced its pH-sensitive aggregation, enabling stabilization of the polyplex outside the cell while still allowing nucleic acid release following cellular entry. The aromatic PEIs were then evaluated for their gene, mRNA, siRNA and 2'O-methyl phosphorothioate oligonucleotide in vitro transfection abilities. The salicylamide-grafted PEI showed to be a reliable carrier for delivering nucleic acids with cytoplasmic activity such as the mRNA and siRNA or nuclear diffusible oligonucleotide. It was then further equipped with polyethyleneglycol (PEG) and the delivery efficiency of the copolymer was tested in vivo for regeneration of dystrophin in the muscle of mdx mouse through a 2'O-methyl phosphorothioate-mediated splicing modulation. Intramuscular administration of polyplexes resulted in dystrophin-positive fibers in a mouse model of Duchenne muscular dystrophy without apparent toxicity. These findings indicate that precise modifications of low molecular weight PEI improve its bio-responsiveness and yield delivery vehicles for nucleic acids of various types in vitro and in vivo.

Published
2017

7. A new method for the formulation of double nanoemulsions

Author

Andrey S. Klymchenko, Shukai Ding, Mériem Er-Rafik, Halina Anton, Wei Yu, Thierry F. Vandamme, Salman Akram, Christophe A. Serra, Nicolas Anton, Institut Charles Sadron (ICS), Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Réseau nanophotonique et optique, Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Matériaux et nanosciences d'Alsace (FMNGE), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Animal et gestion intégrée des risques (UPR AGIRs), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad), Conception et application de molécules bioactives (CAMB), Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Chimie des Systèmes Fonctionnels, Centre National de la Recherche Scientifique (CNRS), Laboratoire de Bioimagerie et Pathologies (LBP), Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Biophotonique et Pharmacologie (CNRS UMR 7213), Laboratoire de Biophotonique et Pharmacologie - UMR 7213 (LBP), Centre National de la Recherche Scientifique (CNRS)-Réseau nanophotonique et optique, Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA)), and Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Aqueous solution, Micrometer scale, Chemical substance, Nanostructure, Materials science, Nanotechnology, 02 engineering and technology, General Chemistry, 010402 general chemistry, 021001 nanoscience & nanotechnology, Condensed Matter Physics, 01 natural sciences, Homogenization (chemistry), 0104 chemical sciences, Microscopy, Emulsion, [SPI.GPROC]Engineering Sciences [physics]/Chemical and Process Engineering, 0210 nano-technology, Science, technology and society
Abstract

Double emulsions are very attractive systems for many reasons; the most important of these are their capacity to encapsulate hydrophilic and lipophilic molecules simultaneously in a single particle and their potentiality to protect fragile hydrophilic molecules from the continuous phase. Double emulsions represent a technology that is widely present down to the micrometer scale; however, double nanoemulsions, with their new potential applications as nanomedicines or diagnosis agents, currently present a significant challenge. In this study, we propose an original two-step approach for the fabrication of double nanoemulsions with a final size below 200 nm. The process consists of the formulation of a primary water-in-oil (w1/O) nanoemulsion by high-pressure homogenization, followed by the re-emulsification of this primary emulsion by a low-energy method to preserve the double nanostructure. Various characterization techniques were undertaken to confirm the double structure and to evaluate the encapsulation efficiency of a small hydrophilic probe in the inner aqueous droplets. Complementary fluorescence confocal and cryo-TEM microscopy experiments were conducted to characterize and confirm the double structure of the double nanoemulsion.

Published
2017

8. Non-linear optical, electrochemical and spectroelectrochemical properties of amphiphilic inner salt porphyrinic systems

Author

Jean-François Nicoud, Karl M. Kadish, Frédéric Bolze, Koen Clays, Lihan Zeng, Nicolas Desbois, Yuanyuan Fang, Claude P. Gros, Yan Cui, Colin Lopez, Hoang Minh Ngo, Griet Depotter, Clément Michelin, Isabelle Ledoux, Institut de Chimie Moléculaire de l'Université de Bourgogne [Dijon] (ICMUB), Université de Bourgogne (UB)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), University of Leuven Department of Chemistry, Université Catholique de Louvain = Catholic University of Louvain (UCL), Department of Chemistry [University of Houston], University of Houston, Laboratoire de Photonique Quantique et Moléculaire (LPQM), École normale supérieure - Cachan (ENS Cachan)-CentraleSupélec-Centre National de la Recherche Scientifique (CNRS), Conception et application de molécules bioactives (CAMB), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), 'Centre National de la Recherche Scientifique' (ICMUB) UMR CNRS 6302, Robert A. Welch Foundation (KMK) E-680, CNRS, the 'Universite de Bourgogne', 'Conseil Regional de Bourgogne' through the 3MIM integrated project ('Marquage de Molecules par les Metaux pour l'Imagerie Medicale'), Institut de Chimie Moléculaire de l'Université de Bourgogne [Dijon] ( ICMUB ), Université de Bourgogne ( UB ) -Centre National de la Recherche Scientifique ( CNRS ), Université Catholique de Louvain ( UCL ), Laboratoire de Photonique Quantique et Moléculaire ( LPQM ), École normale supérieure - Cachan ( ENS Cachan ) -CentraleSupélec-Centre National de la Recherche Scientifique ( CNRS ), Conception et application de molécules bioactives ( CAMB ), and Université de Strasbourg ( UNISTRA ) -Centre National de la Recherche Scientifique ( CNRS )

Subjects
design, Inorganic chemistry, Substituent, amphiphilic inner salt, 02 engineering and technology, 010402 general chemistry, Electrochemistry, Photochemistry, [ CHIM ] Chemical Sciences, electron-accepting moieties, 01 natural sciences, non-linear optical property, chemistry.chemical_compound, Amphiphile, chromophores, [CHIM]Chemical Sciences, Molecule, Chemistry, frequency dispersion, Second-harmonic generation, spectroelectrochemistry, dipolar complexes, General Chemistry, 021001 nanoscience & nanotechnology, Porphyrin, Fluorescence, 0104 chemical sciences, 2nd-harmonic generation, electrochemistry, microscopy, charge-transfer, fluorescence, Absorption (chemistry), hyper-rayleigh scattering, 0210 nano-technology
Abstract

Three zwitterionic meso-substituted A3B- and AB2C-porphyrins containing one sulfonato alkylpyridinium substituent and three or two alkoxy-substituted phenyl groups were synthesized in good yield and fully characterized as to their physicochemical properties by a variety of techniques. This new series of inner salt donor-acceptor meso-substituted porphyrin derivatives were prepared for possible application as amphiphilic probes for membrane insertion in the area of combined second-harmonic and two-photon fluorescence cellular microscopy. To this end, the linear and nonlinear optical properties of the compounds were characterized, together with their electrochemical and spectroelectrochemical properties in non-aqueous media. The neutral design of such molecules enabled us to determine their second order non-linear properties, both by Electric Field Induced Second Harmonic Generation and Hyper–Rayleigh Scattering. Two-photon absorption cross sections of these dyes were also measured by the two-photon induced fluorescence method. The zwitterionic nature of the inner salt results in very specific solvent-dependent redox-properties, which could be rationalized in terms of solvent-dependent ion-pairing. The overall data electrochemical and photophysical data indicates that these new porphyrinic systems should be good probes for membrane potential sensing.

Published
2016

9. Synthèse de nano-vecteurs dérivés des polydiacétylènes pour la co-délivrance d’un ARN interférent et d’un anticancéreux

Author

Ripoll, Manon, Conception et application de molécules bioactives (CAMB), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Université de Strasbourg, Jean-Serge Remy, and Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Thérapie combinatoire, Active targeting, SiRNA, Polydiacetylenes, Anticancer drug, Combination therapy, [CHIM.OTHE]Chemical Sciences/Other, SiARN, Ciblage actif
Abstract

In the nanomedecine field, a new approach consists in developing synthetic vectors able to co-deliver into a cancer cell, an antitumoral drug and siRNAs that target protein(s) involved in MDR. The work described in this manuscript was dedicated to the development of micellar nanovectors for the intracellular co-delivery of these two therapeutic agents. The first part details the synthesis and the formulation of nanometric photopolymerized diacetylenic micelles adapted for the delivery and intracellular release of the siRNA. Then, the encapsulation and delivery properties of these micelles, bearing histidine polar heads have been investigated in vitro and in vivo for the application of combination therapy. Finally, the last part presents the functionalization by electrostatic interaction of these cationic vectors with antibodies, priorly modified by anionic oligonucleotides. This original and versatile system allowed achieving an active targeting of tumoral cells.; En nanomédecine, une nouvelle approche consiste à développer des vecteurs synthétiques pour co-délivrer au sein d’une cellule tumorale, un anticancéreux ainsi qu’un siARN, capable de supprimer l’expression d’une protéine impliquée dans les mécanismes de résistance. Les travaux décrits dans ce manuscrit ont été consacrés à la synthèse de nano-vecteurs micellaires pour la délivrance simultanée de ces deux agents thérapeutiques. Une première partie décrit la synthèse et la formulation de micelles nanométriques diacétyléniques photopolymérisables conçues pour délivrer efficacement un siARN. Les propriétés d’encapsulation et de délivrance de ces micelles ont ensuite été étudiées in vitro et in vivo pour une application en thérapie combinatoire. Enfin, une dernière partie présente la fonctionnalisation par interaction électrostatique de ces vecteurs cationiques avec des anticorps préalablement modifiés par des oligonucléotides anioniques pour réaliser un ciblage actif des cellules tumorales.

Published
2017

10. D-Cateslytin, a new antimicrobial peptide with therapeutic potential

Author

Abdurraouf Zaet, Pauline Dartevelle, Fadoua Daouad, Claire Ehlinger, Fabienne Quilès, Grégory Francius, Christian Boehler, Camille Bergthold, Benoît Frisch, Gilles Prévost, Philippe Lavalle, Francis Schneider, Youssef Haïkel, Marie-Hélène Metz-Boutigue, Céline Marban, Univ Strasbourg, INSERM, UMR S 1121, F-67085 Strasbourg, France, Faculté de chirurgie dentaire - Strasbourg, Université de Strasbourg (UNISTRA), Laboratoire de Chimie Physique et Microbiologie pour l'Environnement (LCPME), Université de Lorraine (UL)-Centre National de la Recherche Scientifique (CNRS), Conception et application de molécules bioactives (CAMB), Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Service de Réanimation Médicale [Strasbourg], Les Hôpitaux Universitaires de Strasbourg (HUS), FRANCIUS, GREGORY, Fédération de Médecine Translationnelle de Strasbourg (FMTS), Institut de Chimie du CNRS (INC)-Université de Lorraine (UL)-Centre National de la Recherche Scientifique (CNRS), Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Hôpital de Hautepierre [Strasbourg], Université de Lorraine (UL)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC), and Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Cell Survival, [SDV]Life Sciences [q-bio], Aucun, lcsh:Medicine, Firmicutes, Microbial Sensitivity Tests, Prevotella intermedia, Permeability, Article, Anti-Infective Agents, Cell Wall, [CHIM] Chemical Sciences, Escherichia coli, Humans, [CHIM]Chemical Sciences, lcsh:Science, Microbial Viability, lcsh:R, Cell Membrane, Drug Synergism, Epithelial Cells, Sciences du Vivant [q-bio]/Microbiologie et Parasitologie, Peptide Fragments, Chromogranin A, lcsh:Q, Caco-2 Cells, Antimicrobial Cationic Peptides
Abstract

International audience; The rise of antimicrobial resistant microorganisms constitutes an increasingly serious threat to global public health. As a consequence, the efficacy of conventional antimicrobials is rapidly declining, threatening the ability of healthcare professionals to cure common infections. Over the last two decades host defense peptides have been identified as an attractive source of new antimicrobials. In the present study, we characterized the antibacterial and mechanistic properties of D-Cateslytin (D-Ctl), a new epipeptide derived from L-Cateslytin, where all L-amino acids were replaced by D-amino acids. We demonstrated that D-Ctl emerges as a potent, safe and robust peptide antimicrobial with undetectable susceptibility to resistance. Using Escherichia coli as a model, we reveal that D-Ctl targets the bacterial cell wall leading to the permeabilization of the membrane and the death of the bacteria. Overall, D-Ctl offers many assets that make it an attractive candidate for the biopharmaceutical development of new antimicrobials either as a single therapy or as a combination therapy as D-Ctl also has the remarkable property to potentiate several antimicrobials of reference such as cefotaxime, amoxicillin and methicillin.

Published
2017

11. Development and evaluation of β-galactosidase-sensitive antibody-drug conjugates

Author

Oleksandr Koniev, Sergii Kolodych, Christian D. Muller, Brigitte Renoux, Sébastien Papot, Alain Wagner, Pauline Poinot, Wojciech Krezel, Jitka Eberova, Chloé Michel, Anthony Ehkirch, Sébastien Delacroix, Sarah Cianférani, Laboratoire de Biomécanique et Mécanique des Chocs (LBMC UMR T9406), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-Institut Français des Sciences et Technologies des Transports, de l'Aménagement et des Réseaux (IFSTTAR), Conception et application de molécules bioactives (CAMB), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Spectrométrie de Masse BioOrganique [Strasbourg] (LSMBO), Département Sciences Analytiques et Interactions Ioniques et Biomoléculaires (DSA-IPHC), Institut Pluridisciplinaire Hubert Curien (IPHC), Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Institut Pluridisciplinaire Hubert Curien (IPHC), Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Institut de Chimie des Milieux et Matériaux de Poitiers (IC2MP), Institut national des sciences de l'Univers (INSU - CNRS)-Centre National de la Recherche Scientifique (CNRS)-Université de Poitiers-Institut de Chimie du CNRS (INC), Institut de génétique et biologie moléculaire et cellulaire (IGBMC), Université Louis Pasteur - Strasbourg I-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Institut Gilbert-Laustriat : Biomolécules, Biotechnologie, Innovation Thérapeutique, Université Louis Pasteur - Strasbourg I-Centre National de la Recherche Scientifique (CNRS), Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)-Institut Pluridisciplinaire Hubert Curien (IPHC), and Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Drug, Antibody-drug conjugate, Immunoconjugates, media_common.quotation_subject, Mice, Nude, Breast Neoplasms, Pharmacology, 010402 general chemistry, Ado-Trastuzumab Emtansine, 01 natural sciences, chemistry.chemical_compound, Antineoplastic Agents, Immunological, Trastuzumab, Cell Line, Tumor, Drug Discovery, medicine, Animals, Humans, [CHIM]Chemical Sciences, Maytansine, ComputingMilieux_MISCELLANEOUS, media_common, Cell Proliferation, 010405 organic chemistry, [CHIM.ORGA]Chemical Sciences/Organic chemistry, Organic Chemistry, Cancer, General Medicine, medicine.disease, beta-Galactosidase, Galactoside, 0104 chemical sciences, 3. Good health, Carcinoma, Ductal, Monomethyl auristatin E, chemistry, Trastuzumab emtansine, Drug delivery, Female, medicine.drug
Abstract

The selective destruction of tumour cells while sparing healthy tissues is one of the main challenges in cancer therapy. Antibody-drug conjugates (ADCs) are arguably the most rapidly expanding class of targeted cancer therapies. Efficient drug conjugation and release technologies are essential for the development of these new therapeutic agents. In response to the ever-increasing demand for efficient drug release systems, we have developed a new class of β-galactosidase-cleavable linkers for ADCs. Within this framework, novel payloads comprising a galactoside linker, the monomethyl auristatin E (MMAE) and cysteine-reactive groups were synthesized, conjugated with trastuzumab and evaluated both in vitro and in vivo. The ADCs with galactoside linkers demonstrated superior therapeutic efficacy in mice compared to the marketed trastuzumab emtansine used for the treatment of breast cancer.

Published
2017

12. Alginate/Chitosan Compact Polyelectrolyte Complexes: A Cell and Bacterial Repellent Material

Author

Fouzia Boulmedais, Béatrice Heurtault, Thida Phoeung, Benoît Frisch, Joseph Hemmerlé, Fabienne Perrin-Schmitt, Dominique Collin, Sylvie Fournel, Michel Rawiso, Andreas Reisch, Philippe Lavalle, Maria Vittoria Spanedda, Emilie Roger, Pierre Schaaf, Angela Mutschler, Régulations Naturelles et Artificielles (ARNA), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Bordeaux Ségalen [Bordeaux 2], Conception et application de molécules bioactives (CAMB), Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Micro et Nanomédecines Translationnelles (MINT), Université d'Angers (UA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Laboratoire d'Hormonologie, Les Hôpitaux Universitaires de Strasbourg (HUS), Immunologie et chimie thérapeutiques (ICT), Cancéropôle du Grand Est-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Biophotonique et Pharmacologie - UMR 7213 (LBP), Centre National de la Recherche Scientifique (CNRS)-Réseau nanophotonique et optique, Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Centre National de la Recherche Scientifique (CNRS), Biomatériaux et Bioingénierie (BB), Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Matériaux et Nanosciences Grand-Est (MNGE), Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Institut de génétique et biologie moléculaire et cellulaire (IGBMC), Université Louis Pasteur - Strasbourg I-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Institut Charles Sadron (ICS), Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Matériaux et Nanosciences Grand-Est (MNGE), Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Réseau nanophotonique et optique, Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Mutschler, Angela, Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA)), Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Matériaux et nanosciences d'Alsace (FMNGE), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Réseau nanophotonique et optique, Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Matériaux et nanosciences d'Alsace (FMNGE), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Université de Strasbourg (UNISTRA)-Matériaux et nanosciences d'Alsace (FMNGE), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM), U1121, and Institut National de la Santé et de la Recherche Médicale (INSERM)

Subjects
[CHIM.POLY] Chemical Sciences/Polymers, Materials science, General Chemical Engineering, Modulus, Salt (chemistry), Alginate chitosan, 02 engineering and technology, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, 010402 general chemistry, Polysaccharide, 01 natural sciences, Chitosan, chemistry.chemical_compound, Polymer chemistry, Materials Chemistry, [SDV.BC] Life Sciences [q-bio]/Cellular Biology, ComputingMilieux_MISCELLANEOUS, chemistry.chemical_classification, [CHIM.MATE] Chemical Sciences/Material chemistry, General Chemistry, [CHIM.MATE]Chemical Sciences/Material chemistry, 021001 nanoscience & nanotechnology, Polyelectrolyte, 0104 chemical sciences, [CHIM.POLY]Chemical Sciences/Polymers, Chemical engineering, chemistry, Homogeneous, Ultracentrifuge, 0210 nano-technology
Abstract

Ultracentrifugated compact polyelectrolyte complexes (uCoPECs) represent a new class of materials that are obtained by ultracentrifugation of solutions of polyanion/polycation complexes in the presence of salt. In the present study, two polysaccharides, chitosan and alginate, were used to form such complexes, thus providing a solid material uniquely composed of polysaccharides. The conditions for obtaining the uCoPEC material were optimized: the optimal salt concentration and polysaccharide concentrations were assessed, and the ultracentrifugation speed proved to be a key parameter to obtain compact and homogeneous materials. The Young's modulus, E, of the material was of the order of 12 MPa, which is the highest E value measured for a uCoPEC. The material contained nanometer-sized crystals of chitosan as indicated by X-ray diffraction. Most strikingly, this material proves to be totally cell- and bacteria-resistant. Immunological tests show that this uCoPEC does not induce any proinflammatory response. T...

Published
2017

13. Synthèse et application d’inducteurs de gènes photo-activables pour le contrôle in vivo de l’expression d’un gène

Author

Goegan, Bastien, Conception et application de molécules bioactives (CAMB), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Université de Strasbourg, Alexandre Specht, and Frédéric Bolze

Subjects
Gene expression system, Photolysis, Tétracycline, [CHIM.ORGA]Chemical Sciences/Organic chemistry, Système d’expression de gène, Two-photon absorption, Photo-removable groups, Tetracycline, Absorption à deux photons, Groupements protecteurs photo-labiles
Abstract

The structural neural network’s is a fundamental process that ensures the proper functioning of the brain. To understand the formation and activity of these networks, we are developing a method which spatio-temporally controlled in vivo, the expression of targeted genes involved in this process at individual neuron cells scale by light. To achieve this in vivo tests, it is necessary to work with methods which are orthogonal to their cellular environment. Photochemical activation by photo-cleavage of an inert biological precursor offers a unique orthogonal way to attain this spatio-temporal control. Therefore, we have recently developed a new family of photoremovable group which are sensitive to two-photon (TP) excitation sensitive, in order to irradiate at favorable wave-lengths for in vivo applications. Moreover, to photo-regulate the expression of genes with high spatial and temporal resolution, we are combining the inducible gene expression system by tetracycline called « Tet-on » system to different photo-activable precursors of tetracycline analogs obtained by hemi-synthesis. All this, should allow us to get an effective system for the in vivo expression of a gene of interest by light excitation in order to photoactivate Kir2.1, a gene that cell autonomously silences the electrical activity of neurons in a subset of cells.; La structuration des réseaux neuronaux est un processus fondamental qui assure le bon fonctionnement du cerveau. Afin de comprendre la formation et l’activité de ces réseaux, nous souhaitons développer une méthode qui permette de contrôler in vivo sous l’action de la lumière l'expression de gènes ciblés impliqués dans ce phénomène,à l’échelle d’une cellule neuronale individuelle. Pour ce faire, nous utilisons une réaction de photo-clivage permettant de libérer de façon contrôlée un inducteur d’expression de gène sous l’action de la lumière, à l’aide de groupements photo-labiles sensibles aux excitations bi-photoniques développés au laboratoire et favorables aux applications in vivo. Afin de photo-réguler l'expression des gènes in vivo et avec un contrôle spatiotemporel élevé, nous combinons le système d’expression de gène inductible par la tétracycline « Tet-on » à une variété de précurseurs photo-activables d’analogues de tétracycline que nous avons synthétisés. Ceci devrait nous permettre de disposer d’un système efficace pour l’expression in vivo d’un gène d’intérêt par excitation lumineuse,et plus précisément dans le but de photo-réguler le gène Kir2.1 impliqué dans la régulation de l’activité électrique des neurones.

Published
2017

14. Synthesis of gene inducers for in vivo photoactivated gene expression

Author

Goegan, Bastien, Conception et application de molécules bioactives (CAMB), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Université de Strasbourg, Alexandre Specht, Frédéric Bolze, and STAR, ABES

Subjects
Gene expression system, Photolysis, Tétracycline, [CHIM.ORGA]Chemical Sciences/Organic chemistry, Système d’expression de gène, Two-photon absorption, Photo-removable groups, Tetracycline, Absorption à deux photons, Groupements protecteurs photo-labiles, [CHIM.ORGA] Chemical Sciences/Organic chemistry
Abstract

The structural neural network’s is a fundamental process that ensures the proper functioning of the brain. To understand the formation and activity of these networks, we are developing a method which spatio-temporally controlled in vivo, the expression of targeted genes involved in this process at individual neuron cells scale by light. To achieve this in vivo tests, it is necessary to work with methods which are orthogonal to their cellular environment. Photochemical activation by photo-cleavage of an inert biological precursor offers a unique orthogonal way to attain this spatio-temporal control. Therefore, we have recently developed a new family of photoremovable group which are sensitive to two-photon (TP) excitation sensitive, in order to irradiate at favorable wave-lengths for in vivo applications. Moreover, to photo-regulate the expression of genes with high spatial and temporal resolution, we are combining the inducible gene expression system by tetracycline called « Tet-on » system to different photo-activable precursors of tetracycline analogs obtained by hemi-synthesis. All this, should allow us to get an effective system for the in vivo expression of a gene of interest by light excitation in order to photoactivate Kir2.1, a gene that cell autonomously silences the electrical activity of neurons in a subset of cells., La structuration des réseaux neuronaux est un processus fondamental qui assure le bon fonctionnement du cerveau. Afin de comprendre la formation et l’activité de ces réseaux, nous souhaitons développer une méthode qui permette de contrôler in vivo sous l’action de la lumière l'expression de gènes ciblés impliqués dans ce phénomène,à l’échelle d’une cellule neuronale individuelle. Pour ce faire, nous utilisons une réaction de photo-clivage permettant de libérer de façon contrôlée un inducteur d’expression de gène sous l’action de la lumière, à l’aide de groupements photo-labiles sensibles aux excitations bi-photoniques développés au laboratoire et favorables aux applications in vivo. Afin de photo-réguler l'expression des gènes in vivo et avec un contrôle spatiotemporel élevé, nous combinons le système d’expression de gène inductible par la tétracycline « Tet-on » à une variété de précurseurs photo-activables d’analogues de tétracycline que nous avons synthétisés. Ceci devrait nous permettre de disposer d’un système efficace pour l’expression in vivo d’un gène d’intérêt par excitation lumineuse,et plus précisément dans le but de photo-réguler le gène Kir2.1 impliqué dans la régulation de l’activité électrique des neurones.

Published
2017

15. Conception et application de nouveaux outils photochimiques pour l’étude des récepteurs canaux P2X

Author

Peverini, Laurie, Conception et application de molécules bioactives (CAMB), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Université de Strasbourg, Alexandre Specht, Thomas Grutter, STAR, ABES, and Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)

Subjects
P2XR, Photochemistry, [CHIM.THER] Chemical Sciences/Medicinal Chemistry, Photochimie, [CHIM.THER]Chemical Sciences/Medicinal Chemistry, Électrophysiologie patch-clamp, Patch-clamp electrophysology
Abstract

P2X receptors are cationic ligand-gated ion channels, activated by extracellular ATP, involved in many physio-pathological roles. Their function is associated with different allosteric states. During this PhD, we have designed three new strategies, spanning photochemical organic synthesis and patch-clamp electrophysiology to elucidate the molecular mechanisms involved in these conformational states and to collect data in order to study structure-function relationships. - Synthesis and application of molecular tweezers, which allows the photo-control of P2X Rand the study of molecular motions - Synthesis and characterization of a photo-cleavable amino acid with the aim of incorporating it into P2XR and doing structure-function relationships - Incorporation of an unnatural amino acid for photo-crosslinking studies. We have been able to probe the molecular mechanism involved in large organic cations permeation of P2XR, to bring into question the dilated state and to identify a physiological cation that can flow through P2XR. We have also designed a photo-cleavable amino acid which could serve in the study of structure-function relationships., Les récepteurs P2X (P2XR), activés par l’ATP, sont impliqués dans des rôles physiopathologiques. Leur fonctionnement est associé à différents états conformationnels. Le projet de thèse a mené à associer la synthèse organique et l’application de molécules photo-activables avec des techniques d’électrophysiologie patch-clamp, pour décortiquer les mouvements moléculaires de ces récepteurs et effectuer des relations structure-fonction, via trois stratégies : - La synthèse et application d’agrafes photo-isomérisables qui permet le photo-contrôle des P2XR et l’étude de mouvements - La synthèse et caractérisation d’un acide aminé (aa) photo-clivable pour étudier les implications de zones sur la fonction des P2XR via une photolyse - L’incorporation d’un aa non naturel dans les P2XR pour étudier des interactions et mouvements via un « photo-pontage ». Nous avons élucidé les mécanismes moléculaires responsables de la perméabilité des P2XR, récusé l’existence de l'état dilaté et identifié un cation organique physiologique pouvant les traverser. Nous avons aussi conçu un acide aminé photo-clivable pouvant mener à des études structure-fonction des P2XR.

Published
2017

16. Stratégies de marquage chimiospécifique et bioorthogonale pour l’analyse métabolomique des rétinoïdes

Author

Thomas, Éric, Conception et application de molécules bioactives (CAMB), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Université de Strasbourg, Alain Wagner, and Wojciech Krezel

Subjects
Retinoids, Bioorthogonal chemistry, Chimie bioorthogonale, Metabolomic, [CHIM.OTHE]Chemical Sciences/Other
Abstract

This work consists of three projects. The first project aims to discover new metabolites of vitamin A. An analog of retinaldehyde, carrying an azide function was synthesized. It would allow to follow its fate in vivo. The second project consisted in the development of a probe allowing the analysis of all the aldehyde metabolites in a sample. The probe provides sensitivity gain in LS-MS². An analysis of its biodistribution has been done, and showed the ATPP probe is distributed after an intraperitoneal injection. Concerning the third project, a homobifunctional coupling reagent "thiol-to-thiol" has been developed. The coupling products showed excellent plasma stability. The reagent was first successfully applied to the coupling of small molecules and then to the coupling of a modified oligonucleotide and a peptide.; Ce travail est composé de trois projets. Le premier projet a pour objectif de découvrir de nouveaux métabolites de la vitamine A. Il a consisté en la synthèse d’un analogue du rétinaldéhyde, portant une fonction azoture et permettant de suivre son devenir in vivo. Le second projet a consisté en l’élaboration de la sonde ATPP permettant l’analyse de l’ensemble des métabolites aldéhydiques d’un échantillon. La sonde permet un gain de sensibilité en LS-MS². Une analyse de sa biodistribution a été faite, et montre que la sonde ATPP, après injection intrapéritonéale, est distribuée in vivo. Concernant le troisième projet, un réactif de couplage homobifonctionnel « thiol-thiol » a été élaboré. Les produits du couplage ont montré une excellente stabilité plasmatique. Le réactif a d’abord été appliqué avec succès au couplage de petites molécules, puis au couplage d’un oligonucléotide modifié et d’un peptide.

Published
2017

17. Development of innovative liposome-based constructs for non-invasive cancer immunotherapy in humans

Author

Saliba, Hanadi, Conception et application de molécules bioactives (CAMB), Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Université de Strasbourg, Sylvie Fournel, Soulaima Chamat, Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), and STAR, ABES

Subjects
Liposome, [SDV.CAN] Life Sciences [q-bio]/Cancer, Tumor-specific vaccine, Vaccin antitumoral, [SDV.CAN]Life Sciences [q-bio]/Cancer, Souris humanisée, [SDV.IMM.IMM]Life Sciences [q-bio]/Immunology/Immunotherapy, [SDV.IMM.IMM] Life Sciences [q-bio]/Immunology/Immunotherapy, Humanized mouse, Transcutaneous route, Voie transcutanée
Abstract

A vaccine administration route and the preclinical model in which it is evaluated are major factors that contribute to its success in humans. In this context, the discovery that the transcutaneous (TC) route induces a powerful immune response has made the TC tumor-specific vaccination a promising strategy. Further evaluation of candidate vaccines in a humanized mouse model (Hu-SPL-NSG), more predictive of the human response, is also needed. The objective of this thesis is to (i) optimize liposomal constructs incorporating peptides and a TLR agonist for the TC pathway and (ii) evaluate their immunogenicity in the Hu-SPL-NSG model. Thus, we have varied the nature of the TLR agonist and the deformability of the liposomal vesicle, and have added a dendritic cell targeting molecule. Immunogenicity of these formulations by the TC route was then evaluated in mice. Finally, we tested the ability of a model liposomal construct to induce a cellular and humoral response in the Hu-SPL-NSG model.All of this work provided a first proof of concept on the feasibility of TC tumor-specific vaccination by liposomes and its applicability in humans., La voie d’administration d’un vaccin et le modèle préclinique dans le lequel il est évalué sont des facteurs majeurs qui contribuent à son succès chez l’homme. Dans ce contexte, la découverte que la voie transcutanée (TC) induit une réponse immunitaire puissante a fait de la vaccination antitumorale TC une stratégie prometteuse. Une évaluation complémentaire du candidat vaccin dans un modèle de souris humanisée (Hu-SPL-NSG), plus prédictif de la réponse humaine, est aussi nécessaire. L’objectif de cette thèse est i) d'optimiser des constructions liposomiques peptidiques incorporant un agoniste de TLR pour la voie TC et ii) d’évaluer leur immunogénicité dans le modèle Hu-SPL-NSG. Ainsi, nous avons fait varier la nature de l’agoniste de TLR et la déformabilité de la vésicule liposomique, et avons rajouté une molécule de ciblage des cellules dendritiques. L’immunogenicité de ces formulations par voie TC a ensuite été évaluée chez la souris. Enfin, nous avons testé la capacité d’une construction liposomique modèle à induire une réponse cellulaire et humorale dans le modèle Hu-SPL-NSG. L’ensemble de ces travaux a fourni une première preuve de concept sur la faisabilité de la vaccination antitumorale TC par des liposomes et de son applicabilité chez l’homme.

Published
2017

18. Conjugués d'anticorps : approche intégrative pour une bioconjugaison plus sélective, stable et contrôlable

Author

Dovgan, Igor, Conception et application de molécules bioactives (CAMB), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Université de Strasbourg, Alain Wagner, and Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Mono-functionalisation of proteins, [CHIM.ORGA]Chemical Sciences/Organic chemistry, Lysine, Bioconjugation, Antibody-drug conjugates, Conjugués d'anticorps, Lysine conjugation, Arginine, Antibody-oligonucleotide conjugates, Arginine conjugation
Abstract

Within the last decade, antibodies conjugated to cytotoxic drugs or oligonucleotides have gained a great attention in scientific community owing to the unique properties of the antibodies, such as their long circulation time in serum and high selectivity against their target. For instance, antibody conjugates (ACs) are increasingly applied for targeted cancer therapy or bioimaging. Consequently, the development of reliable methodologies for ACs preparation is currently of high demand. However, the controllable conjugation and preparation of ACs with defined structure are still challenging due to high excess and variety of reactive groups in antibody structure, which are accessible for conjugation. Moreover, current linker technologies are based on the maleimide-thiol reaction, yielding adducts, which are unstable during circulation in blood.This work is focused on chemical approaches for the reliable antibody functionalisation, which enable the preparation of stable ACs with well-defined payload to antibody ratios. The first part is devoted to design and development of maleimide-dioxane reagents as self-hydrolysable and serum-stable alternative to classical maleimide chemistry. The second part is dedicated to a screening approach for evaluation of residue-selective functionalities in reactions with an antibody using high resolution native mass spectrometry. Finally, in the third part the reader is introduced with a novel technology, which enables efficient preparation of stable ACs with a defined degree of conjugation and particularly mono-functionalisation of antibodies.; Au cours de la dernière décennie, les anticorps conjugués à des médicaments cytotoxiques ou des oligonucléotides ont acquis une grande attention dans la communauté scientifique en raison des propriétés uniques des anticorps, tels que leur long temps de circulation dans le sérum et leur sélectivité élevée par rapport à leur cible. Par exemple, les conjugués d'anticorps (ACs) sont de plus en plus appliqués en thérapie ciblée contre le cancer ou en bioimagerie. Par conséquent, le développement de méthodologies fiables pour la préparation des AC est actuellement en pleine expansion. Cependant, la conjugaison et la préparation contrôlables des ACs avec une structure définie rencontrent encore de nombreux obstacles en raison de l'excès élevé et de la variété des groupes réactifs dans la structure des anticorps, qui sont accessibles pour la conjugaison. En outre, les technologies de liaison actuelles sont basées sur la réaction de maléimide-thiol, produisant des adduits, qui sont instables dans le sang. Ce travail se concentre sur les approches chimiques pour la fonctionnalisation fiable des anticorps, qui permettent la préparation d'ACs stables présentant un ratio anticorps/principe actif bien défini. La première partie est consacrée à la conception et au développement du réactif maléimide-dioxane, solution auto-hydrolysable et stable dans le sérum, comme alternative à la chimie classique du maléimide. La deuxième partie est consacrée à l'évaluation de la réactivité sélective des différents acides aminés portés par les anticorps par spectrométrie de masse native à haute résolution. Finalement, une nouvelle technologie permettant d’obtenir des ACs stables avec un ratio anticorps/principe actif contrôlé est présentée au lecteur dans une 3ème partie.

Published
2017

19. Massively parallel and multiparameter titration of biochemical assays with droplet microfluidics

Author

Yannick Rondelez, Jean-François Bartolo, Valérie Taly, Nicolas Bredeche, Shu Okumura, Elia Henry, Teruo Fujii, Rémi Sieskind, Nathanael Aubert-Kato, Alexandre Baccouche, Anthony J. Genot, TALY, Valerie, Retour Post-Doctorant - Nano Puce digitial a ADN - - DigiNANO2013 - ANR-13-PDOC-0001 - PDOC - VALID, Laboratory for Integrated Micro Mechatronics Systems (LIMMS), The University of Tokyo (UTokyo)-Centre National de la Recherche Scientifique (CNRS), Earth-Life Science Institute [Tokyo] (ELSI), Tokyo Institute of Technology [Tokyo] (TITECH), Institute of Industrial Science (IIS), The University of Tokyo (UTokyo), Gulliver (UMR 7083), Ecole Superieure de Physique et de Chimie Industrielles de la Ville de Paris (ESPCI Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Department of Information Science [Tokyo, Japan], Ochanomizu University, Institut supérieur technologique Montplaisir (ISTM), Conception et application de molécules bioactives (CAMB), Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Médecine Personnalisée, Pharmacogénomique, Optimisation Thérapeutique (MEPPOT - U1147), Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM), Equipe labellisée Ligue contre le Cancer, Y.R. acknowledges support from the Japan Society for the Promotion of Science (JSPS) through a Grant-in-Aid for Scientific Research on Innovative Areas ‘Synthetic Biology for Comprehension of Biomolecular Networks’ (project number 23119001). A.J.G. acknowledges support from the ANR (ANR-13-PDOC-0001) and the JSPS (postdoctoral fellowship).J.-F.B. was supported by a PhD fellowship from Region Alsace. N.B. acknowledges support from the PHC Sakura program (project number 34171WG). A.B. and N.A.-K. were supported by the ELSI Origins Network (EON), which is supported by a grant from the John Templeton Foundation., ANR-13-PDOC-0001,DigiNANO,Nano Puce digitial a ADN(2013), Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Ecole Superieure de Physique et de Chimie Industrielles de la Ville de Paris (ESPCI Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL), Centre National de la Recherche Scientifique (CNRS)-The University of Tokyo (UTokyo), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), The University of Tokyo-Centre National de la Recherche Scientifique (CNRS), The University of Tokyo, Gulliver, ESPCI ParisTech-Centre National de la Recherche Scientifique (CNRS), Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Equipe Labellisée par la Ligue Nationale contre le Cancer, and LNCC

Subjects
[SDV.BIO]Life Sciences [q-bio]/Biotechnology, [SDV]Life Sciences [q-bio], Microfluidics, 02 engineering and technology, Bioinformatics, Barcode, 01 natural sciences, General Biochemistry, Genetics and Molecular Biology, law.invention, Chemical kinetics, Surface-Active Agents, law, Fluorescence microscope, Image Processing, Computer-Assisted, Massively parallel, Fluorescent Dyes, Chemistry, 010401 analytical chemistry, Titrimetry, Equipment Design, Microfluidic Analytical Techniques, 021001 nanoscience & nanotechnology, Fluorescence, 0104 chemical sciences, [SDV.BIO] Life Sciences [q-bio]/Biotechnology, [SDV] Life Sciences [q-bio], Reagent, Titration, Biological Assay, 0210 nano-technology, Biological system
Abstract

International audience; Biochemical systems in which multiple components take part in a given reaction are of increasing interest. Because the interactions between these different components are complex and difficult to predict from basic reaction kinetics, it is important to test for the effect of variations in the concentration for each reagent in a combinatorial manner. For example, in PCR, an increase in the concentration of primers initially increases template amplification, but large amounts of primers result in primer-dimer by-products that inhibit the amplification of the template. Manual titration of biochemical mixtures rapidly becomes costly and laborious, forcing scientists to settle for suboptimal concentrations. Here we present a droplet-based microfluidics platform for mapping of the concentration space of up to three reaction components followed by detection with a fluorescent readout. The concentration of each reaction component is read through its internal standard (barcode), which is fluorescent but chemically orthogonal. We describe in detail the workflow, which comprises the following: (i) production of the microfluidics chips, (ii) preparation of the biochemical mixes, (iii) their mixing and compartmentalization into water-in-oil emulsion droplets via microfluidics, (iv) incubation and imaging of the fluorescent barcode and reporter signals by fluorescence microscopy and (v) image processing and data analysis. We also provide recommendations for choosing the appropriate fluorescent markers, programming the pressure profiles and analyzing the generated data. Overall, this platform allows a researcher with a few weeks of training to acquire ∼10,000 data points (in a 1D, 2D or 3D concentration space) over the course of a day from as little as 100-1,000 μl of reaction mix.

Published
2017

20. Functionalizing Nanoemulsions with Carboxylates: Impact on the Biodistribution and Pharmacokinetics in Mice

Author

Mayeul Collot, Sidy Mouhamed Dieng, Caroline Bouillot, Thierry F. Vandamme, Andrey S. Klymchenko, Christophe A. Serra, Mériem Er-Rafik, Mohamed F. Attia, Nicolas Anton, Marc Schmutz, Conception et application de molécules bioactives (CAMB), Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Chimie des Systèmes Fonctionnels, Centre National de la Recherche Scientifique (CNRS), Laboratoire de Bioimagerie et Pathologies (LBP), Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Biophotonique et Pharmacologie (CNRS UMR 7213), Laboratoire de Biophotonique et Pharmacologie - UMR 7213 (LBP), Centre National de la Recherche Scientifique (CNRS)-Réseau nanophotonique et optique, Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA)), Institut Charles Sadron (ICS), Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Réseau nanophotonique et optique, Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Matériaux et nanosciences d'Alsace (FMNGE), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Animal et gestion intégrée des risques (UPR AGIRs), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad), and Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Biodistribution, Polymers and Plastics, Bioengineering, Nanotechnology, 02 engineering and technology, 010402 general chemistry, 01 natural sciences, Biomaterials, Mice, Pharmacokinetics, Lipid droplet, Materials Chemistry, Animals, [SPI.GPROC]Engineering Sciences [physics]/Chemical and Process Engineering, Maleic Anhydrides, chemistry.chemical_classification, Drug Carriers, Chemistry, Polymer, 021001 nanoscience & nanotechnology, Lipids, 0104 chemical sciences, Targeted drug delivery, Biophysics, Surface modification, Emulsions, Nanocarriers, 0210 nano-technology, Drug carrier, Biotechnology
Abstract

Efficiency of drug administration is related to the inhibition of adverse effects, and can be improved by drug targeting through lipid nanocarriers encapsulation. Targeting technology generally goes along with the nanocarrier functionalization that can be surface modification and/or ligand grafting. The great advantage of nanoemulsions is their loading capability and the possibilities to encapsulate several entities in a single droplet, however, the decoration of the lipid droplets with strongly anchored reactive functions is challenging. This study proposes a reliable and innovative method to functionalize lipid droplets, based on the lipophilic polymer poly(maleic anhydride-alt-1-octadecene), solubilized in the droplet core, and able to hydrolyze at the oil/water interface. Interfacial chemistry and physicochemical properties of nanodroplets are characterized. In vitro studies reveal that the presence of carboxylates at interface has a strong impact on the interactions with cells, as the internalization of functionalized droplets is much higher than control ones. This difference is confirmed with longitudinal computed tomography studies in mice after i.v. administration, strongly impacting the pharmacokinetics and biodistributions. This work establishes the proof-of-concept of a new method for functionalizing lipid droplets and demonstrates that surface modification can have a significant impact on their interaction with cells, pharmacokinetics, and biodistribution.

Published
2017

21. Solubilization of α-galactosylceramide in aqueous medium: Impact on Natural Killer T cell activation and antitumor responses

Author

Christelle Faveeuw, François Trottein, Maria Vittoria Spanedda, Marcella De Giorgi, Benoît Frisch, Neila Chekkat, Béatrice Heurtault, Sylvie Fournel, Christophe Ehret, Jean-Sébastien Thomann, Line Bourel-Bonnet, Didier Betbeder, Elodie Macho-Fernandez, Centre d’Infection et d’Immunité de Lille - INSERM U 1019 - UMR 9017 - UMR 8204 (CIIL), Centre National de la Recherche Scientifique (CNRS)-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille)-Université de Lille-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Pasteur de Lille, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP), Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille), Conception et application de molécules bioactives (CAMB), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Lille Inflammation Research International Center - U 995 (LIRIC), Institut Pasteur de Lille, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille), Université d'Artois (UA), This work was supported by Institut National du Cancer (INCa) grant INCA_6699, Ligue contre le Cancer, Alsace contre le Cancer, the Centre National de la Recherche Scientifique and the Université de Strasbourg., Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille)-Centre National de la Recherche Scientifique (CNRS), Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), and Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille-Institut Pasteur de Lille

Subjects
0301 basic medicine, Male, Lung Neoplasms, medicine.medical_treatment, Pharmaceutical Science, chemical and pharmacologic phenomena, [SDV.CAN]Life Sciences [q-bio]/Cancer, Antineoplastic Agents, Galactosylceramides, Pharmacology, Lymphocyte Activation, 03 medical and health sciences, Mice, 0302 clinical medicine, Immune system, In vivo, Cell Line, Tumor, medicine, Animals, Antigen-presenting cell, Water-soluble α-GalCer, Micelles, Cancer, Liposome, Drug Carriers, Chemistry, α-GalCer formulations, Natural killer T cell, In vitro, 3. Good health, carbohydrates (lipids), Mice, Inbred C57BL, 030104 developmental biology, Cytokine, Biochemistry, [SDV.IMM.IA]Life Sciences [q-bio]/Immunology/Adaptive immunology, 030220 oncology & carcinogenesis, iNKT, Liposomes, Natural Killer T-Cells, lipids (amino acids, peptides, and proteins), [SDV.IMM.VAC]Life Sciences [q-bio]/Immunology/Vaccinology, Drug carrier
Abstract

International audience; The potent antitumor effect of α-galactosylceramide (α-GalCer) is based on its recognition by invariant Natural Killer T cells (iNKT) after its capture and presentation by antigen presenting cells including dendritic cells (DCs). Synthetic α-GalCer has already been tested in advanced cancer patients but no or only moderate clinical responses were obtained.To optimize α-GalCer efficacy, we have postulated that alternative formulations impacting its molecular organization in aqueous medium could modify DC uptake and iNKT-based immune responses. To this end, we have developed two strategies: (1) the formulation of α-GalCer in non-cationic liposomes and (2) the synthesis of a water-soluble α-GalCer analogue by anchoring a polyethyleneglycol moiety on its sugar head. The biological activities of these new preparations were compared to that induced by the classically used Polysorbate 20 α-GalCer micelles. Both formulations retained their uptake by DCs and activated iNKT cells both in vitro and in vivo. Despite a lower cytokine production, the formulations induced a potent immune response able to control lung murine carcinoma. In conclusion, it is possible to increase α-GalCer solubility in aqueous solution without limiting its antitumor properties.

Published
2017

22. Identification of autoreactive B cells with labeled nucleosomes

Author

Cécile Seifert, A. Guffroy, Thierry Martin, Anne-M Knapp, Anne-S Korganow, Jean-L Pasquali, Jean-D Fauny, Pauline Soulas-Sprauel, Vincent Gies, Alain Wagner, Hélène Dumortier, Immuno-Rhumatologie Moléculaire, Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM), Conception et application de molécules bioactives (CAMB), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Chimie des Systèmes Fonctionnels, Centre National de la Recherche Scientifique (CNRS), Immunopathologie et chimie thérapeutique (ICT), Institut de biologie moléculaire et cellulaire (IBMC), Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), and Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)

Subjects
0301 basic medicine, Science, Transgene, Autoimmunity, [SDV.BC]Life Sciences [q-bio]/Cellular Biology, medicine.disease_cause, Autoantigens, Article, Cell Line, Flow cytometry, Mice, 03 medical and health sciences, 0302 clinical medicine, Sciences du Vivant [q-bio]/Biologie cellulaire, medicine, Animals, Humans, Lupus Erythematosus, Systemic, B cell, Autoantibodies, B-Lymphocytes, Multidisciplinary, Lupus erythematosus, Staining and Labeling, biology, medicine.diagnostic_test, Autoantibody, Flow Cytometry, medicine.disease, Molecular biology, Nucleosomes, 3. Good health, Cell biology, 030104 developmental biology, Histone, medicine.anatomical_structure, Cell culture, biology.protein, Medicine, Female, Biomarkers, 030215 immunology
Abstract

The pathogenesis of autoimmune diseases has not been completely elucidated yet, and only a few specific treatments have been developed so far. In autoimmune diseases mediated by pathogenic autoantibodies, such as systemic lupus erythematosus, the specific detection and analysis of autoreactive B cells is crucial for a better understanding of the physiopathology. Biological characterization of these cells may help to define new therapeutic targets. Very few techniques allowing the precise detection of autoreactive B cells have been described so far. Herein we propose a new flow cytometry technique for specific detection of anti-nucleosome B cells, which secrete autoantibodies in systemic lupus erythematosus, using labeled nucleosomes. We produced different fluorochrome-labeled nucleosomes, characterized them, and finally tested them in flow cytometry. Nucleosomes labeled via the cysteines present in H3 histone specifically bind to autoreactive B cells in the anti-DNA transgenic B6.56R mice model. The present work validates the use of fluorochrome-labeled nucleosomes via cysteines to identify anti-nucleosome B cells and offers new opportunities for the description of autoreactive B cell phenotype.

Published
2017

23. Novel 3D in vitro models based on multicellular tumor spheroids to test anticancer drugs and drug delivery vehicles

Author

Akasov, Roman, Conception et application de molécules bioactives (CAMB), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Université de Strasbourg, M.M. Shemyakin-Yu. A. Ovchinnikova Institute of bioorganic chemistry (Moscow), Thierry Vandamme, Elena Markvicheva, STAR, ABES, and Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Tumeurs, Curcumin, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, Doxorubicine, Anticancer drugs, Cyclic RGD-peptides, [SDV.BIO] Life Sciences [q-bio]/Biotechnology, Microréservoirs, Microcontainers, Nano-emulsions, Sphéroïdes, Doxorubicin, Peptides RGD cycliques, Nano-émulsions, Spheroids, Curcumine, Micelles, Principes actifs anticancéreux, Tumors
Abstract

Multicellular tumor spheroids (MTS) are a promising tool in tumor biology. The aim of the Thesis was to develop a novel highly reproducible technique for MTS formation, and to demonstrate the availability of these spheroids as 3D in vitro model to test anticancer drugs and drug delivery vehicles. Cell self-assembly effect induced by an addition of cyclic RGD-peptides directly to monolayer cultures was studied for 16 cell lines of various origin. Cyclo-RGDfK peptide and its modification with triphenylphosphonium cation (TPP) were found to induce spheroid formation. The spheroids were used as a model to evaluate the cytotoxicity of antitumor drugs (doxorubicin, curcumin, temozolomide) and a number of nano- and micro- formulations (microcontainers, nano-emulsions and micelles)., Les sphéroïdes multicellulaires tumoraux (SMT) constituent un outil prometteur dans le domaine de l’étude biologique des tumeurs. Le but de la thèse était de développer une technique de la formation de SMT et de démontrer la disponibilité de ces sphéroïdes comme modèle in vitro 3D pour tester l’efficacité de principes actifs anticancéreux ainsi que celle de formulations de délivrance de médicaments. L'effet d’auto-assemblage de cellules induit par une addition des peptides RGD cycliques a été étudié pour 16 lignées cellulaires de différentes origines. Le peptide cyclique RGDfK et sa modification avec le cation triphenylphosphonium (TPP) ont permis de mettre en évidence l’induction de formation de sphéroïdes. Les sphéroïdes ont été employés comme modèles pour évaluer la cytotoxicité de principes actifs antitumoraux (doxorubicine, curcumine, temozolomide) et un certain nombre de formulations nano- et micrométriques (microréservoirs, nano-émulsions et micelles).

Published
2017

24. In situ targeted activation of an anticancer agent using ultrasound-triggered release of composite droplets

Author

Kolodych, Sergii, Michel, Chloé, DELACROIX, Sébastien, Koniev, Oleksandr, Ehkirch, Anthony, Eberova, Jitka, Cianferani, Sarah, Krezel, Wojciech, Poinot, Pauline, Muller, Christian, Wagner, Alain, Bezagu, Marine, Clarhaut, Jonathan, Renoux, Brigitte, Monti, Fabrice, Tanter, Mickaël, Tabeling, Patrick, Cossy, Janine, Couture, Olivier, Papot, Sébastien, Arseniyadis, Stellios, Laboratoire d'Analyse du Mouvement, Institut national de podologie, Conception et application de molécules bioactives (CAMB), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Institut Pluridisciplinaire Hubert Curien (IPHC), Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Institut de génétique et biologie moléculaire et cellulaire (IGBMC), Université Louis Pasteur - Strasbourg I-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Ecole Nationale Vétérinaire, Agroalimentaire et de l'alimentation Nantes-Atlantique (ONIRIS), Cooltech Applications, Cooltech, Ecole Superieure de Physique et de Chimie Industrielles de la Ville de Paris (ESPCI Paris), Université Paris sciences et lettres (PSL), Institut de physiologie et biologie cellulaires (IPBC), Université de Poitiers-Centre National de la Recherche Scientifique (CNRS), Institut de Chimie des Milieux et Matériaux de Poitiers (IC2MP), Institut national des sciences de l'Univers (INSU - CNRS)-Centre National de la Recherche Scientifique (CNRS)-Université de Poitiers-Institut de Chimie du CNRS (INC), Gulliver (UMR 7083), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Institut Langevin - Ondes et Images (UMR7587) (IL), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP), Laboratoire de Physique Statistique de l'ENS (LPS), Université Paris Diderot - Paris 7 (UPD7)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS)-Fédération de recherche du Département de physique de l'Ecole Normale Supérieure - ENS Paris (FRDPENS), Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL), Laboratoire de Chimie Oraganique, Institut de Mathématiques de Bourgogne [Dijon] (IMB), Centre National de la Recherche Scientifique (CNRS)-Université de Franche-Comté (UFC), Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université Bourgogne Franche-Comté [COMUE] (UBFC)-Université de Bourgogne (UB), School of Biological and Chemical Sciences, Queen Mary University of London, Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Université de Strasbourg (UNISTRA)-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS), Université de Poitiers-Institut national des sciences de l'Univers (INSU - CNRS)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS), Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de Physique Nucléaire et de Physique des Particules du CNRS (IN2P3)-Centre National de la Recherche Scientifique (CNRS), École nationale vétérinaire, agroalimentaire et de l'alimentation Nantes-Atlantique (ONIRIS), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Fédération de recherche du Département de physique de l'Ecole Normale Supérieure - ENS Paris (FRDPENS), École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Paris (ENS-PSL), and Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS)

Subjects
0301 basic medicine, Drug, Cell Survival, media_common.quotation_subject, Context (language use), Antineoplastic Agents, Pharmacology, 03 medical and health sciences, 0302 clinical medicine, Therapeutic index, Drug Delivery Systems, Glucuronides, Pharmacokinetics, Cell Line, Tumor, Neoplasms, Drug Discovery, Distribution (pharmacology), Humans, Prodrugs, Ultrasonics, Cytotoxicity, ComputingMilieux_MISCELLANEOUS, media_common, Glucuronidase, Chemistry, Organic Chemistry, General Medicine, [CHIM.CATA]Chemical Sciences/Catalysis, Prodrug, 3. Good health, 030104 developmental biology, 030220 oncology & carcinogenesis, Cancer cell, Biophysics, Oligopeptides
Abstract

The efficiency of a drug is usually highly dependent on the way it is administered or delivered. As such, targeted-therapy, which requires conceiving drug-delivery vehicles that will change their state from a relatively stable structure with a very slow leak-rate to an unstable structure with a fast release, clearly improves the pharmacokinetics, the absorption, the distribution, the metabolism and the therapeutic index of a given drug. In this context, we have developed a particularly effective double stimuli-responsive drug-delivery method allowing an ultrasound-induced release of a monomethylauristatin E-glucuronide prodrug and its subsequent activation by a β-glucuronidase. This led to an increase of cytotoxicity of about 80% on cancer cells.

Published
2017

25. Targeting the tumour microenvironment with an enzyme-responsive drug delivery system for the efficient therapy of breast and pancreatic cancers

Author

Sébastien Papot, Balkis Eddhif, Alain Le Pape, Jérôme Alsarraf, Isabelle Tranoy-Opalinski, Brigitte Renoux, Thibaut Legigan, Pauline Poinot, Florian Raes, Elodie Péraudeau, Stéphanie Lerondel, Jonathan Clarhaut, Sergii Kolodych, Oleksandr Koniev, Institut de Chimie des Milieux et Matériaux de Poitiers (IC2MP), Institut national des sciences de l'Univers (INSU - CNRS)-Centre National de la Recherche Scientifique (CNRS)-Université de Poitiers-Institut de Chimie du CNRS (INC), Transgenèse et archivage d'animaux modèles (TAAM), Centre National de la Recherche Scientifique (CNRS), Centre hospitalier universitaire de Poitiers (CHU Poitiers), Ecole Nationale Vétérinaire, Agroalimentaire et de l'alimentation Nantes-Atlantique (ONIRIS), Conception et application de molécules bioactives (CAMB), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Transgénèse et Archivage d'Animaux Modèles - Centre d'Imagerie du Petit Animal (TAAM-CIPA), Institut de physiologie et biologie cellulaires (IPBC), and Université de Poitiers-Centre National de la Recherche Scientifique (CNRS)

Subjects
Drug, Cancer chemotherapy, media_common.quotation_subject, 02 engineering and technology, 010402 general chemistry, 01 natural sciences, Monomethylauristatin E, Extracellular, Medicine, ComputingMilieux_MISCELLANEOUS, media_common, chemistry.chemical_classification, business.industry, Albumin, General Chemistry, [CHIM.CATA]Chemical Sciences/Catalysis, 021001 nanoscience & nanotechnology, 0104 chemical sciences, 3. Good health, Enzyme, chemistry, Immunology, Drug delivery, Cancer research, 0210 nano-technology, business
Abstract

The development of novel therapeutic strategies allowing the destruction of tumour cells while sparing healthy tissues is one of the main challenges of cancer chemotherapy. Here, we report on the design and antitumour activity of a low-molecular-weight drug delivery system programmed for the selective release of the potent monomethylauristatin E in the tumour microenvironment of solid tumours. After intravenous administration, this compound binds covalently to plasmatic albumin through Michael addition, thereby enabling its passive accumulation in tumours where extracellular β-glucuronidase initiates the selective release of the drug. This targeting device produces outstanding therapeutic efficacy on orthotopic triple-negative mammary and pancreatic tumours in mice (50% and 33% of mice with the respective tumours cured), leading to impressive reduction or even disappearance of tumours without inducing side effects.

Published
2017

26. Extrusion-sphéronisation de produits pharmaceutiques : système de délivrance des principes actifs peu solubles par voie orale

Author

Nguyen, Thi Trinh Lan, STAR, ABES, Conception et application de molécules bioactives (CAMB), Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Université de Strasbourg, Thierry Vandamme, and Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Principe actif faiblement soluble, Minigranule, Water-insoluble drug, Improved solubility, Extrusion-spheronization, Folic acid, [CHIM.THER] Chemical Sciences/Medicinal Chemistry, Kétoprofène, [CHIM.THER]Chemical Sciences/Medicinal Chemistry, Amélioration de la solubilité, Lit fluidisé, Pellet, Ketoprofen, Extrusion-sphéronisation, Fluid bed coater, Acide folique
Abstract

Improvement in dissolution of poorly soluble drugs has many challenges.In this thesis, an extrusion-spheronization process was thoroughly studied forimproving dissolubility of drug with nano-emulsion formulation.The aim of the thesis work is to describe the properties and manufacturing processes ofpellets to increase the solubility of poorly soluble active ingredients in water and thus improvetheir bioavailability when administered orally: folic acid (water-insoluble vitamin) andketoprofen (Non-steroidal anti-inflammatory, having anti-inflammatory, analgesic andantipyretic action, class II in the Biopharmaceutical Classification System).This study describes the preparation by extrusion-spheronization, characterisation andin vitro dissolution study of folic acid and ketoprofen pellets. Ketoprofen pellets coated withAcryl-EZE®, Advantia® Performance in a fluid-bed minicoater. The results of the tests showedthe feasibility of the preparation of enteric-coated pellets containing a NSAID and that bycoating the multiparticulate system with either 17.5% Acryl-EZE® 93A92545 or with 12%Advantia® Performance 190024HA49 weight gain, an enteric release of the drug from thepellets can be obtained. The results of dissolution testing indicated that in acidic media, entericfilm coating resulted in a delay in the release of the drug, while no delay was observed in pH6.8 buffer media., L'amélioration de la dissolution des médicaments peu solubles présente de nombreux défis.Dans cette thèse, un procédé d'extrusion-sphéronisation a été étudié en profondeur pour améliorer la dissolubilité du médicament avec une formulation de nano-émulsion. Le but du travail de thèse est de décrire les propriétés et les procédés de fabrication de minigranules permettant d'augmenter la solubilité des principes actifs peu solubles dans l'eau et donc d‘améliorer leur biodisponibilité lors de l'administration par voie orale, pour deux modèles de molécules différentes qui sont l‘acide folique (vitamine peu soluble dans l'eau) et le kétoprofène (anti-inflammatoire non stéroïdien qui présente une solubilité limitée dans les fluides gastriques à cause de son pKa (classe II dans le système de classification biopharmaceutique – BCS, ayant une action anti-inflammatoire, antalgique et antipyrétique). Cette étude décrit la préparation par extrusion-sphéronisation, caractérisation et étude de dissolution in vitro d'acide folique et de pastilles de kétoprofène revêtues de Acryl-EZE®, Advantia® Performance dans un minicoatère à lit fluidisé. Les résultats des essais ont montré la faisabilité de la préparation de pastilles enrobées entériques contenant un AINS et que, en revêtant le système multiparticulaire avec Acryl-EZE® 93A92545 et Advantia® Performance190024HA49 à un gain pondéral de 17,5%, 12,0%, respectivement, du médicament à partir des pastilles peuvent être obtenus. Les résultats des essais de dissolution ont indiqué que dans un milieu acide, le revêtement de film a entraîné un retard dans la libération du médicament, alors qu'aucun retard n'a été observé dans un milieu tampon à pH 6,8.

Published
2017

27. 3D in vitro co-culture models based on normal cells and tumor spheroids formed by cyclic RGD-peptide induced cell self-assembly

Author

Elena Markvicheva, Sergey V. Burov, Emmanuel Guedon, Daria Zaytseva-Zotova, Isabelle Chevalot, Anastasia Gileva, Roman Akasov, Conception et application de molécules bioactives (CAMB), Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Shemyakin & Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences [Moscow] (RAS), Laboratoire Réactions et Génie des Procédés (LRGP), Université de Lorraine (UL)-Centre National de la Recherche Scientifique (CNRS), and Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)

Subjects
0301 basic medicine, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, [SDV]Life Sciences [q-bio], Tumor spheroid, Cell, Bioengineering, Nanotechnology, Applied Microbiology and Biotechnology, Peptides, Cyclic, law.invention, 03 medical and health sciences, Mice, Confocal microscopy, law, Cell Line, Tumor, Spheroids, Cellular, medicine, Animals, ComputingMilieux_MISCELLANEOUS, Cell Nucleus, Microscopy, Confocal, Chemistry, Spheroid, General Medicine, Fibroblasts, In vitro, Coculture Techniques, 030104 developmental biology, medicine.anatomical_structure, embryonic structures, Drug delivery, Biophysics, Self-assembly, Co-Culture, Biotechnology
Abstract

To design novel 3D in vitro co-culture models based on the RGD-peptide-induced cell self-assembly technique. Multicellular spheroids from M-3 murine melanoma cells and L-929 murine fibroblasts were obtained directly from monolayer culture by addition of culture medium containing cyclic RGD-peptide. To reach reproducible architecture of co-culture spheroids, two novel 3D in vitro models with well pronounced core–shell structure from tumor spheroids and single mouse fibroblasts were developed based on this approach. The first was a combination of a RGD-peptide platform with the liquid overlay technique with further co-cultivation for 1–2 days. The second allowed co-culture spheroids to generate within polyelectrolyte microcapsules by cultivation for 2 weeks. M-3 cells (a core) and L-929 fibroblasts (a shell) were easily distinguished by confocal microscopy due to cell staining with DiO and DiI dyes, respectively. The 3D co-culture spheroids are proposed as a tool in tumor biology to study cell–cell interactions as well as for testing novel anticancer drugs and drug delivery vehicles.

Published
2017

28. Non-invasive quantitative imaging of hepatocellular carcinoma growth in mice by micro-CT using liver-targeted iodinated nano-emulsions

Author

Mohamed F. Attia, Hugues Jacobs, Tania Sorg, Alexandru Parlog, Eric Robinet, Isabelle Goncalves, Marie Wattenhofer-Donzé, Ghina Bou About, Thierry F. Vandamme, Nicolas Anton, Laboratoire de Conception et Application de Molécules Bioactives, Centre National de la Recherche Scientifique (CNRS), Conception et application de molécules bioactives (CAMB), Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA), Laboratoire de Conception et Application de Molécules Bioactives (UMR 7199 CNRS/Université de Strasbourg), CNRS (UMR 7199), Laboratoire de Chimie des Systèmes Fonctionnels, Institut de génétique et biologie moléculaire et cellulaire (IGBMC), Université Louis Pasteur - Strasbourg I-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Centre for Integrative Biology - CBI (Inserm U964 - CNRS UMR7104 - IGBMC), Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institute of genetics and molecular and cellular biology, Institut Clinique de la Souris, Institut de Recherche sur les Maladies Virales et Hépatiques (IVH), Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), and Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)

Subjects
0301 basic medicine, Quantitative imaging, Carcinoma, Hepatocellular, Halogenation, Nano emulsion, lcsh:Medicine, Sciences du Vivant [q-bio]/Médecine humaine et pathologie, Article, 03 medical and health sciences, Mice, 0302 clinical medicine, Cell Line, Tumor, Carcinoma, medicine, Animals, Humans, Nanotechnology, lcsh:Science, Cell Proliferation, Ultrasonography, Multidisciplinary, medicine.diagnostic_test, business.industry, Non invasive, Liver Neoplasms, lcsh:R, Cancer, Magnetic resonance imaging, X-Ray Microtomography, medicine.disease, 3. Good health, 030104 developmental biology, medicine.anatomical_structure, Cell Transformation, Neoplastic, Liver, 030220 oncology & carcinogenesis, Hepatocellular carcinoma, Hepatocyte, Emulsions, Female, lcsh:Q, Nuclear medicine, business, [SDV.MHEP]Life Sciences [q-bio]/Human health and pathology
Abstract

Hepatocellular carcinoma (HCC) is the only cancer for which non-invasive diagnosis is recognized by international guidelines. Contrast agent free ultrasound imaging, computed tomography (CT) and/or magnetic resonance imaging are techniques used for early detection and confirmation. Clinical evidence depicts that CT is 30% less precise as compared to MRI for detection of small tumors. In our work, we have reported some novel tools that can enhance the sensitivity and precision of CT applied to preclinical research (micro-CT). Our system, containing non-toxic nano-droplets loaded with iodine has high contrasting properties, liver and hepatocyte specificity and strong liver persistence. Micro-CT was performed on HCC model implanted in nude mice by intrahepatic injection. Contrast agent was administrated intravenously. This method allows an unprecedented high precision of detection, quantitative measurement of tumor volume and quantitative follow-up of the tumor development.

Published
2017

29. Therapeutical modulation of plasmacytoid dendritic cells in experimental arthritis

Author

Birgit Niederreiter, Jean Sibilia, Antonia Puchner, Gernot Schabbauer, Philippe Kastner, Thierry F. Vandamme, Seiamak Bahram, Christopher G. Mueller, Victoria Saferding, Vincent Flacher, Stephan Blüml, Martin Holcmann, Philippe Georgel, Benjamin Voisin, Peggy Kirstetter, Susan Chan, Ghada Alsaleh, Ramzi Nehmar, Alexandre Mariotte, Immuno-Rhumatologie Moléculaire, Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM), Immunopathologie et chimie thérapeutique (ICT), Institut de biologie moléculaire et cellulaire (IBMC), Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Chimie des Systèmes Fonctionnels, Centre National de la Recherche Scientifique (CNRS), Conception et application de molécules bioactives (CAMB), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Centre for Integrative Biology - CBI (Inserm U964 - CNRS UMR7104 - IGBMC), Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC), Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Institut de génétique et biologie moléculaire et cellulaire (IGBMC), Université Louis Pasteur - Strasbourg I-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), univOAK, Archive ouverte, Fédération de Médecine Translationnelle de Strasbourg (FMTS), Université de Strasbourg (UNISTRA), Centre de Recherche d’Immunologie et d’Hématologie [Strasbourg], Centre de recherche du médicament Medalis - Drug Discovery Center [LabEx], Medizinische Universität Wien = Medical University of Vienna, Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA), and Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)

Subjects
0301 basic medicine, rheumatoid arthritis, [SDV]Life Sciences [q-bio], medicine.medical_treatment, Arthritis, Imiquimod, Arthritis, Rheumatoid, Mice, 0302 clinical medicine, Interferon, Immunology and Allergy, Medicine, Mice, Knockout, [SDV.MHEP] Life Sciences [q-bio]/Human health and pathology, Membrane Glycoproteins, Reverse Transcriptase Polymerase Chain Reaction, hemic and immune systems, Flow Cytometry, 3. Good health, Cytokine, plasmacytoid dendritic cells, Rheumatoid arthritis, Interferon Type I, Aminoquinolines, Cytokines, Tumor necrosis factor alpha, medicine.symptom, medicine.drug, Immunology, Inflammation, Enzyme-Linked Immunosorbent Assay, Mice, Transgenic, Sciences du Vivant [q-bio]/Médecine humaine et pathologie, 03 medical and health sciences, Ikaros Transcription Factor, Rheumatology, Adjuvants, Immunologic, Animals, Humans, 030203 arthritis & rheumatology, business.industry, Tumor Necrosis Factor-alpha, Gene Expression Profiling, Dendritic Cells, medicine.disease, Arthritis, Experimental, imiquimod, Disease Models, Animal, 030104 developmental biology, Toll-Like Receptor 7, business, Interferon type I, [SDV.MHEP]Life Sciences [q-bio]/Human health and pathology
Abstract

International audience; Objective: The role of plasmacytoid dendritic cells (PDCs) and type I interferons (IFNs) in rheumatoid arthritis (RA) remains a subject of controversy. This study was undertaken to explore the contribution of PDCs and type I IFNs to RA pathogenesis using various animal models of PDC depletion and to monitor the effect of localized PDC recruitment and activation on joint inflammation and bone damage.Methods: Mice with K/BxN serum-induced arthritis, collagen-induced arthritis, and human tumor necrosis factor transgene insertion were studied. Symptoms were evaluated by visual scoring, quantification of paw swelling, determination of cytokine levels by enzyme-linked immunosorbent assay, and histologic analysis. Imiquimod-dependent therapeutic effects were monitored by transcriptome analysis (using quantitative reverse transcriptase-polymerase chain reaction) and flow cytometric analysis of the periarticular tissue.Results: PDC-deficient mice showed exacerbation of inflammatory and arthritis symptoms after arthritogenic serum transfer. In contrast, enhancing PDC recruitment and activation to arthritic joints by topical application of the Toll-like receptor 7 (TLR-7) agonist imiquimod significantly ameliorated arthritis in various mouse models. Imiquimod induced an IFN signature and led to reduced infiltration of inflammatory cells.Conclusion: The therapeutic effects of imiquimod on joint inflammation and bone destruction are dependent on TLR-7 sensing by PDCs and type I IFN signaling. Our findings indicate that local recruitment and activation of PDCs represents an attractive therapeutic opportunity for RA patients.

Published
2017

30. The absorption enhancer sodium deoxycholate promotes high gene transfer in skeletal muscles

Author

Daniel Scherman, Laetitia van Wittenberghe, Pascal Bigey, Nelly El Shafey, Christian Leborgne, Antoine Kichler, Debborah Alimi-Guez, Approches génétiques intégrées et nouvelles thérapies pour les maladies rares (INTEGRARE), Université d'Évry-Val-d'Essonne (UEVE)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Généthon-École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL), Unité de Technologies Chimiques et Biologiques pour la Santé (UTCBS - UM 4 (UMR 8258 / U1022)), Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC), Conception et application de molécules bioactives (CAMB), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université d'Évry-Val-d'Essonne (UEVE)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Généthon, Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), École Pratique des Hautes Études (EPHE), and Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université Paris Descartes - Paris 5 (UPD5)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)

Subjects
0301 basic medicine, [SDV.BIO]Life Sciences [q-bio]/Biotechnology, Cell Membrane Permeability, Cell Survival, Transgene, [SDV]Life Sciences [q-bio], Pharmaceutical Science, Biology, Muscle disorder, Gene delivery, Injections, Intramuscular, Cell Line, 03 medical and health sciences, 0302 clinical medicine, Genes, Reporter, Luciferases, Firefly, medicine, Animals, Enhancer, Luciferases, Muscle, Skeletal, Reporter gene, Mice, Inbred BALB C, Gene Transfer Techniques, Skeletal muscle, DNA, Alkaline Phosphatase, 3. Good health, 030104 developmental biology, medicine.anatomical_structure, Biochemistry, Lac Operon, Naked DNA, 030220 oncology & carcinogenesis, Alkaline phosphatase, Female, Deoxycholic Acid
Abstract

International audience; Gene delivery to skeletal muscle is a promising strategy for the treatment of muscle disorders and for the systemic secretion of therapeutic proteins. In addition, muscle is an attractive target tissue because it is easily accessible. However, very few synthetic vectors proved capable of surpassing naked DNA mediated muscle gene transfer. In fact, only neutral copolymers, in particular poloxamers, demonstrated capacities to increase transgene expression in skeletal muscles. Here, we studied in vitro and in vivo behaviour of different bile salts. We report that sodium deoxycholate (DOC) and derivatives thereof increase after intramuscular injection by more than 100-fold the levels of the reporter gene luciferase compared to naked DNA. Using a LacZ expression cassette, we found that more than 20% of the muscle fibers expressed the reporter gene. Prolonged expression of a secreted reporter gene derived from a natural murine alkaline phosphatase enzyme could be documented. Altogether, our results demonstrate that bile salts belong to the most efficient chemicals identified so far for skeletal muscle gene transfer. Importantly, since these compounds are naturally found in the body, there is no risk of immune response against them and in addition several bile salts are already used in human medicine. Bile salt mediated muscle gene transfer may thus have broad applications in gene therapy.

Published
2016

31. Iodinated nano-emulsions as contrast agents for preclinical X-ray imaging: Impact of the free surfactants on the pharmacokinetics

Author

Stéphanie Briançon, Hatem Fessi, Thierry F. Vandamme, Nicolas Anton, François Hallouard, Xiang Li, Laboratoire d'automatique et de génie des procédés (LAGEP), Université Claude Bernard Lyon 1 (UCBL), Université de Lyon-Université de Lyon-École Supérieure Chimie Physique Électronique de Lyon-Centre National de la Recherche Scientifique (CNRS), Conception et application de molécules bioactives (CAMB), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), and GARRIGUES, Olivier

Subjects
Biodistribution, [SPI.GPROC] Engineering Sciences [physics]/Chemical and Process Engineering, Contrast Media, Pharmaceutical Science, chemistry.chemical_element, Nanotechnology, 02 engineering and technology, Kidney, 010402 general chemistry, Iodine, 01 natural sciences, Suspension (chemistry), law.invention, Mice, Surface-Active Agents, Pulmonary surfactant, Pharmacokinetics, law, Phase (matter), Animals, Tissue Distribution, [SPI.GPROC]Engineering Sciences [physics]/Chemical and Process Engineering, Filtration, Chromatography, ACL, technology, industry, and agriculture, Iodized Oil, General Medicine, [SDV.SP]Life Sciences [q-bio]/Pharmaceutical sciences, 021001 nanoscience & nanotechnology, 0104 chemical sciences, [SDV.SP] Life Sciences [q-bio]/Pharmaceutical sciences, Liver, chemistry, Drug Design, Nanoparticles, Emulsions, Tomography, X-Ray Computed, 0210 nano-technology, Preclinical imaging, Half-Life, Biotechnology
Abstract

This study presents new important aspects in the design of contrast agents for X-ray preclinical imaging. The first one is a new simple formulation of long circulating contrast agents, formulated from a commercial iodinated oil, and resulting in CT contrast agents containing more than twice the iodine concentration commercial contrast agents. The second point is a methodological aspect, utilizing tangential filtration for reducing the residual surfactants in the bulk phase and serving as well for concentrating droplets (and iodine) in the suspension. The last point is a more general aspect regarding the influence of the free surfactant on the pharmacokinetics and biodistribution of the nano-emulsion droplets on mice. We showed that cross-flow filtration is efficient for concentrating the droplets and reducing the concentration of free surfactant from 10 wt.% to 1 wt.%, without any changes in the nano-emulsion droplet morphologies or surface properties. We also showed that the presence of free surfactant has a significant impact on the elimination way of the nano-emulsion droplets, shared between liver and kidneys. The purified nano-emulsions are preferentially eliminated by the kidneys in contrast to raw nano-emulsions, predominantly eliminated by the liver. In practice, for two similar suspensions, half-life decreases from 4.1 ± 1.10 h to 2.5 ± 0.77 h before and after purification. Since the design and development of long circulating systems are critical in numerous domains, and not for preclinical CT imaging, this study presents important results in that field, taken under a formulation and technical point of view.

Published
2013

32. N-Heterocyclic Carbene–Polyethylenimine Platinum Complexes with Potent in Vitro and in Vivo Antitumor Efficacy

Author

Gilles Guichard, Stéphane Bellemin-Laponnaz, Marion Decossas, Olivier Lambert, Sylvie Fournel, Riccardo Rubbiani, Georges Dahm, Edith Chardon, Benoît Frisch, Gilles Gasser, May Wantz, Justine Sitz, Neila Chekkat, Institut de Physique et Chimie des Matériaux de Strasbourg (IPCMS), Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Matériaux et nanosciences d'Alsace (FMNGE), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Réseau nanophotonique et optique, Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA), Immunologie et chimie thérapeutiques (ICT), Cancéropôle du Grand Est-Centre National de la Recherche Scientifique (CNRS), Ecole Nationale Vétérinaire Agroalimentaire et de l'Alimentation Nantes Atlantique (ONIRIS), Conception et application de molécules bioactives (CAMB), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Ecole Nationale Supérieure de Chimie de Paris - Chimie ParisTech-PSL (ENSCP), Université Paris sciences et lettres (PSL), Université Louis Pasteur - Laboratoire Decomet (UMR 7177-LC3), Université Louis Pasteur - Strasbourg I-Commencez à saisir le nom d'une tutelle, University of Zurich, and Fournel, Sylvie

Subjects
10120 Department of Chemistry, Programmed cell death, Organoplatinum Compounds, Biomedical Engineering, 3003 Pharmaceutical Science, Pharmaceutical Science, chemistry.chemical_element, 2204 Biomedical Engineering, Bioengineering, Antineoplastic Agents, [SDV.CAN]Life Sciences [q-bio]/Cancer, [CHIM.THER]Chemical Sciences/Medicinal Chemistry, Pharmacology, Mitochondrion, 010402 general chemistry, 01 natural sciences, chemistry.chemical_compound, Mice, In vivo, Cell Line, Tumor, 540 Chemistry, medicine, Animals, Humans, Polyethyleneimine, ComputingMilieux_MISCELLANEOUS, Polyethylenimine, 1502 Bioengineering, 010405 organic chemistry, Chemistry, Organic Chemistry, Biological Transport, Xenograft Model Antitumor Assays, In vitro, 3. Good health, 0104 chemical sciences, Oxaliplatin, Mitochondria, 3004 Pharmacology, [CHIM.POLY]Chemical Sciences/Polymers, Cancer research, 1305 Biotechnology, Platinum, Biotechnology, medicine.drug, Conjugate, 1605 Organic Chemistry
Abstract

The current interest for platinum N-heterocyclic carbene complexes in cancer research stems from their impressive toxicity reported against a range of different human cancer cells. To date, the demonstration of their in vivo efficacy relative to that of established platinum-based drugs has not been specifically addressed. Here, we introduce an innovative approach to increase the NHC-Pt complex potency whereby multiple NHC-Pt(II) complexes are coordinated along a polyethylenimine polymer (PEI) chain. We show that such NHC-Pt(II)-PEI conjugates induce human cancer cell death in vitro and in vivo in a xenograft mouse model with no observable side effects in contrast to oxaliplatin. Additional studies indicate nucleus and mitochondria targeting and suggest various mechanisms of action compared to classical platinum-based anticancer drugs.

Published
2016

33. Fluorescent Tobacco mosaic virus-derived bio-nanoparticles for intravital two-photon imaging

Author

Boudewijn van der Sanden, Annette Niehl, Sonia Boscá, Florence Appaix, Jean-François Nicoud, Frédéric Bolze, Manfred Heinlein, Institut de biologie moléculaire des plantes (IBMP), Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA), Conception et application de molécules bioactives (CAMB), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Chimie des Systèmes Fonctionnels, Centre National de la Recherche Scientifique (CNRS), and univOAK, Archive ouverte

Subjects
0301 basic medicine, Plant virus, 02 engineering and technology, Plant Science, Biology, lcsh:Plant culture, Blood–brain barrier, 03 medical and health sciences, Two-photon excitation microscopy, Tobacco mosaic virus, medicine, Fluorescence microscope, [SDV.BV]Life Sciences [q-bio]/Vegetal Biology, [SDV.BV] Life Sciences [q-bio]/Vegetal Biology, lcsh:SB1-1110, two-photon microscopy, Original Research, viral nanoparticles, 021001 nanoscience & nanotechnology, Virology, Fluorescence, 030104 developmental biology, medicine.anatomical_structure, Biophysics, intravital imaging, Nanocarriers, 0210 nano-technology, Preclinical imaging
Abstract

Multi-photon intravital imaging has become a powerful tool to investigate the healthy and diseased brain vasculature in living animals. Although agents for multi-photon fluorescence microscopy of the microvasculature are available, issues related to stability, bioavailability, toxicity, cost or chemical adaptability remain to be solved. In particular, there is a need for highly fluorescent dyes linked to particles that do not cross the blood brain barrier (BBB) in brain diseases like tumor or stroke to estimate the functional blood supply. Plant virus particles possess a number of distinct advantages over other particles, the most important being the multi-valency of chemically addressable sites on the particle surface. This multi-valency, together with biological compatibility and inert nature, makes plant viruses ideal carriers for in vivo imaging agents. Here, we show that the well-known Tobacco mosaic virus is a suitable nanocarrier for two-photon dyes and for intravital imaging of the mouse brain vasculature.

Published
2016

34. Enantioselective ethylation of aldehydes using a regenerable polymer-supported N-picolylvalinol tridentate ligand

Author

Vinciane Kelsen, Philippe C. Gros, Philippe Pierrat, Laboratoire de physique et chimie des nano-objets (LPCNO), Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut National des Sciences Appliquées (INSA)-Université de Toulouse (UT)-Institut de Chimie de Toulouse (ICT), Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT)-Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT)-Institut de Recherche sur les Systèmes Atomiques et Moléculaires Complexes (IRSAMC), Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Université de Toulouse (UT)-Centre National de la Recherche Scientifique (CNRS)-Centre National de la Recherche Scientifique (CNRS), Conception et application de molécules bioactives (CAMB), Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Institut Français de Recherche pour l'Exploitation de la Mer (IFREMER), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Institut de Chimie de Toulouse (ICT-FR 2599), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Institut de Recherche pour le Développement (IRD)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie du CNRS (INC)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie du CNRS (INC)-Institut de Recherche sur les Systèmes Atomiques et Moléculaires Complexes (IRSAMC), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Centre National de la Recherche Scientifique (CNRS)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), and Institut Français de Recherche pour l'Exploitation de la Mer - Atlantique (IFREMER Atlantique)

Subjects
Tridentate ligand, [CHIM.ORGA]Chemical Sciences/Organic chemistry, 010405 organic chemistry, Organic Chemistry, Chiral ligand, Enantioselective synthesis, [CHIM.CATA]Chemical Sciences/Catalysis, Diethylzinc, 010402 general chemistry, 01 natural sciences, Biochemistry, 0104 chemical sciences, Catalysis, chemistry.chemical_compound, chemistry, Drug Discovery, Pyridine, Organic chemistry, Enantiomer, ComputingMilieux_MISCELLANEOUS, Polymer supported
Abstract

A supported pyridine-based tridentate chiral ligand has been prepared and evaluated for enantioselective addition of diethylzinc to aldehydes. The catalyst allowed the preparation of various chiral alcohols with the R configuration in good yields and enantiomeric excesses (up to 93%) and was found reusable.

Published
2007

35. Molecular structure and function of P2X receptors

Author

Thomas Grutter, Kate Dunning, Thierry Chataigneau, Chloé Habermacher, Laboratoire de Chimie des Systèmes Fonctionnels, Centre National de la Recherche Scientifique (CNRS), Conception et application de molécules bioactives (CAMB), and Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)

Subjects
0301 basic medicine, Protein Conformation, Allosteric regulation, Context (language use), Molecular engineering, 03 medical and health sciences, Cellular and Molecular Neuroscience, Adenosine Triphosphate, Allosteric Regulation, Protein Domains, Animals, Humans, Receptor, Ion channel, Pharmacology, Binding Sites, Chemistry, Molecular biophysics, Purinergic receptor, Combinatorial chemistry, Protein Structure, Tertiary, Optogenetics, 030104 developmental biology, Receptors, Purinergic P2X, [SDV.NEU]Life Sciences [q-bio]/Neurons and Cognition [q-bio.NC], Neuroscience, Ion Channel Gating, Function (biology), Protein Binding
Abstract

ATP-gated P2X receptors are trimeric ion channels selective to cations. Recent progress in the molecular biophysics of these channels enables a better understanding of their function. In particular, data obtained from biochemical, electrophysiogical and molecular engineering in the light of recent X-ray structures now allow delineation of the principles of ligand binding, channel opening and allosteric modulation. However, although a picture emerges as to how ATP triggers channel opening, there are a number of intriguing questions that remain to be answered, in particular how the pore itself opens in response to ATP and how the intracellular domain, for which structural information is limited, moves during activation. In this review, we provide a summary of functional studies in the context of the post-structure era, aiming to clarify our understanding of the way in which P2X receptors function in response to ATP binding, as well as the mechanism by which allosteric modulators are able to regulate receptor function. This article is part of the Special Issue entitled 'Purines in Neurodegeneration and Neuroregeneration'.

Published
2015

36. Human recombinant thiamine triphosphatase: purification, secondary structure and catalytic properties

Author

Bernard Lakaye, Benaissa El Moualij, Ridha Aichour, Laurence Lins, Ilca Margineanu, Pierre Wins, Willy Zorzi, Thierry Grisar, Bernard Coumans, Lucien Bettendorff, Luc Lebeau, Alexander F Makarchikov, Séverine Roland, Center for Cellular and Molecular Neurobiology, Universi de Lge, Chimie et Biologie des Membranes et des Nanoobjets (CBMN), Université de Bordeaux (UB)-École Nationale d'Ingénieurs des Travaux Agricoles - Bordeaux (ENITAB)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Conception et application de molécules bioactives (CAMB), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Institute of Human Histology-CRPP (CRPP), and Université de Liège

Subjects
DNA, Complementary, Cations, Divalent, [SDV]Life Sciences [q-bio], Thiamin-Triphosphatase, Regulatory site, Thiamine Triphosphate, Biochemistry, Catalysis, Protein Structure, Secondary, Substrate Specificity, 03 medical and health sciences, Enzyme activator, chemistry.chemical_compound, Adenosine Triphosphate, Thiamine triphosphatase, Cerebellum, Diethyl Pyrocarbonate, Enzyme Stability, Escherichia coli, Humans, Cloning, Molecular, Binding site, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology, chemistry.chemical_classification, 0303 health sciences, Binding Sites, Molecular Structure, biology, Chemistry, 030302 biochemistry & molecular biology, Active site, Cell Biology, Recombinant Proteins, Enzyme Activation, Enzyme, biology.protein, Thiamine triphosphate
Abstract

Thiamine triphosphate (ThTP) is found in most living organisms and it may act as a phosphate donor for protein phosphorylation. We have recently cloned the cDNA coding for a highly specific mammalian 25 kDa thiamine triphosphatase (ThTPase; EC 3.6.1.28). As the enzyme has a high catalytic efficiency and no sequence homology with known phosphohydrolases, it was worth investigating its structure and catalytic properties. For this purpose, we expressed the untagged recombinant human ThTPase (hThTPase) in E. coli, produced the protein on a large scale and purified it to homogeneity. Its kinetic properties were similar to those of the genuine human enzyme, indicating that the recombinant hThTPase is completely functional. Mg2+ ions were required for activity and Ca2+ inhibited the enzyme by competition with Mg2+. With ATP as substrate, the catalytic efficiency was 10(-4)-fold lower than with ThTP, confirming the nearly absolute specificity of the 25 kDa ThTPase for ThTP. The activity was maximum at pH 8.5 and very low at pH 6.0. Zn2+ ions were inhibitory at micromolar concentrations at pH 8.0 but activated at pH 6.0. Kinetic analysis suggests an activator site for Mg2+ and a separate regulatory site for Zn2+. The effects of group-specific reagents such as Woodward's reagent K and diethylpyrocarbonate suggest that at least one carboxyl group in the active site is essential for catalysis, while a positively charged amino group may be involved in substrate binding. The secondary structure of the enzyme, as determined by Fourier-transform infrared spectroscopy, was predominantly beta-sheet and alpha-helix.

Published
2004

37. Performance of Pyridylthiourea-Polyethylenimine Polyplex for siRNA-Mediated Liver Cancer Therapy in Cell Monolayer, Spheroid, and Tumor Xenograft Models

Author

Francine Gossé, Mathieu Gonçalves, Florent Meyer, Benoît Frisch, Cendrine Seguin, Guy Zuber, Jean Baptiste Gossart, Etienne Pascal, Emilie Heuillard, Eric Robinet, Interaction virus-hôte et maladies du foie, Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM), Conception et application de molécules bioactives (CAMB), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Institut de Chimie des Substances Naturelles (ICSN), Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC), and Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)

Subjects
0301 basic medicine, Small interfering RNA, [SDV.CAN]Life Sciences [q-bio]/Cancer, 02 engineering and technology, delivery system, Biology, liver cancer, polo‐like kinase, 03 medical and health sciences, chemistry.chemical_compound, In vivo, RNA interference, medicine, ComputingMilieux_MISCELLANEOUS, Gene knockdown, Polyethylenimine, Full Paper, Spheroid, Cancer, Sciences du Vivant [q-bio]/Biotechnologies, [SDV.IMM.IMM]Life Sciences [q-bio]/Immunology/Immunotherapy, Full Papers, 021001 nanoscience & nanotechnology, medicine.disease, Molecular biology, polyethylenimine, 3. Good health, [CHIM.POLY]Chemical Sciences/Polymers, 030104 developmental biology, chemistry, Tumor progression, siRNA, Cancer research, 0210 nano-technology
Abstract

Medical application of siRNAs relies on methods for delivering nucleic acids into the cytosol. Synthetic carriers, which assemble with nucleic acids into delivery systems, show promises for cancer therapy but efficiency remains to be improved. In here, the effectiveness of pyridylthiourea‐polyethylenimine (πPEI), a siRNA carrier that favors both polyplex disassembly and endosome rupture upon sensing the acidic endosomal environment, in 3 experimental models of hepatocellular cancer is tested. The πPEI‐assisted delivery of a siRNA targeting the polo‐like kinase 1 into Huh‐7 monolayer produces a 90% cell death via a demonstrated RNA interference mechanism. Incubation of polyplex with Huh‐7 spheroids leads to siRNA delivery into the superficial first cell layer and a 60% reduction in spheroid growth compared to untreated controls. Administration of polyplexes into mice bearing subcutaneous implanted Huh‐7Luc tumors results in a reduced tumor progression, similar to the one observed in the spheroid model. Altogether, these results support the in vivo use of synthetic and dedicated polymers for increasing siRNA‐mediated gene knockdown, and their clinical promise in cancer therapeutics.

Published
2017

38. Polycationic Pillar[5]arene Derivatives: Interaction with DNA and Biological Applications

Author

Michel Holler, Jean-François Nierengarten, Marc Nothisen, David Sigwalt, Iwona Nierengarten, Thomas Biellmann, Jean-Serge Remy, Laboratoire d'innovation moléculaire et applications (LIMA), Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Conception et application de molécules bioactives (CAMB), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Institut de Chimie de Strasbourg, and Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Dendrimers, [SDV.OT]Life Sciences [q-bio]/Other [q-bio.OT], Stereochemistry, Nanoparticle, [CHIM.THER]Chemical Sciences/Medicinal Chemistry, 010402 general chemistry, Cleavage (embryo), Transfection, 01 natural sciences, Catalysis, chemistry.chemical_compound, Dynamic light scattering, Dendrimer, Polyamines, Humans, ComputingMilieux_MISCELLANEOUS, Gel electrophoresis, 010405 organic chemistry, [CHIM.ORGA]Chemical Sciences/Organic chemistry, Organic Chemistry, General Chemistry, DNA, Combinatorial chemistry, Polyelectrolytes, 0104 chemical sciences, Quaternary Ammonium Compounds, chemistry, Click chemistry, Nanoparticles, Amine gas treating, Click Chemistry, sense organs, Calixarenes, HeLa Cells, Plasmids
Abstract

Dendritic pillar[5]arene derivatives have been efficiently prepared by grafting dendrons with peripheral Boc-protected amine subunits onto a preconstructed pillar[5]arene scaffold. Upon cleavage of the Boc-protected groups, water-soluble pillar[5]arene derivatives with 20 (13) and 40 (14) peripheral ammonium groups have been obtained. The capability of these compounds to form stable nanoparticles with plasmid DNA has been demonstrated by gel electrophoresis, transmission electron microscopy (TEM), and dynamic light scattering (DLS) investigations. Transfection efficiencies of the self-assembled 13/pCMV-Luc and 14/pCMV-Luc polyplexes have been evaluated in vitro with HeLa cells. The transfection efficiencies found for both compounds are good, and pillar[5]arenes 13 and 14 show very low toxicity if any.

Published
2013

39. Allosteric signaling and dynamics of the clamshell-like NMDA receptor GluN1 N-terminal domain

Author

Zhu, Shujia, Stroebel, David, Yao, C Andrea, TALY, Antoine, Paoletti, Pierre, Hénin, Jérôme, Institut de biologie de l'ENS Paris (IBENS), Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Département de Biologie - ENS Paris, École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Conception et application de molécules bioactives (CAMB), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Section Neuroscience, Institut de biologie de l'ENS Paris (UMR 8197/1024) (IBENS), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Département de Biologie - ENS Paris, Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM)-École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-Institut National de la Santé et de la Recherche Médicale (INSERM), and Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)

Subjects
[SDV.BBM.BP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biophysics, [SDV.BBM.BP] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Biophysics, ComputingMilieux_MISCELLANEOUS
Abstract

International audience; no abstract

Published
2013

40. Direct Electrochemistry of Cytochrome bo Oxidase at a series of Gold Nanoparticles-Modified Electrodes

Author

Melin, Frederic, Meyer, Thomas, Lankiang, Styven, Choi, Sylvia K., Gennis, Robert B., Blanck, Christian, Schmutz, Marc, Hellwig, Petra, Laboratoire de spectroscopie vibrationnelle et électrochimie des biomolécules, Institut de Chimie de Strasbourg, Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)-Institut de Chimie du CNRS (INC), University of Illinois at Urbana-Champaign [Urbana], University of Illinois System, Institut Charles Sadron (ICS), Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Réseau nanophotonique et optique, Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Matériaux et nanosciences d'Alsace (FMNGE), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Matériaux et Nanosciences Grand-Est (MNGE), Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Réseau nanophotonique et optique, Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), and univOAK, Archive ouverte

Subjects
Électrochimie, [CHIM.CATA] Chemical Sciences/Catalysis, Spectroscopie, Spectroélectrochimie, [CHIM.CATA]Chemical Sciences/Catalysis, Chimie Physique
Abstract

PMID: 23335854 PMCID: PMC3546826; New membrane-protein based electrodes were prepared incorporating cytochrome bo(3) from E. coli and gold nanoparticles. Direct electron transfer between the electrode and the immobilized enzymes was achieved, resulting in an electrocatalytic activity in presence of O(2). The size of the gold nanoparticles was shown to be important and smaller particles were shown to reduce the overpotential of the process.

Published
2012

41. Key role for respiratory CD103(+) dendritic cells, IFN-γ, and IL-17 in protection against Streptococcus pneumoniae infection in response to α-galactosylceramide

Author

Laurye Van Maele, Jean-Claude Sirard, Hélène Léger, Bernard Ryffel, Christelle Faveeuw, Stoyan Ivanov, Elodie Macho Fernandez, Natalia Muñoz Wolf, Analía Rial, Christophe Paget, Arndt Benecke, Joelle Renneson, Benoît Frisch, François Trottein, José A. Chabalgoity, Isabelle Maillet, Josette Fontaine, Centre d’Infection et d’Immunité de Lille - INSERM U 1019 - UMR 9017 - UMR 8204 (CIIL), Institut Pasteur de Lille, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Lille-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille)-Centre National de la Recherche Scientifique (CNRS), Immunologie et Embryologie Moléculaires (IEM), Université d'Orléans (UO)-Centre National de la Recherche Scientifique (CNRS), Conception et application de molécules bioactives (CAMB), Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Universidad de la República [Montevideo] (UDELAR), Institut des Hautes Études Scientifiques (IHES), IHES, Institut de Recherche Interdisciplinaire [Villeneuve d'Ascq] (IRI), Université de Lille, Sciences et Technologies-Université de Lille, Droit et Santé-Centre National de la Recherche Scientifique (CNRS), Institut National de la Santé et de la Recherche Médicale, CNRS,University of Lille Nord de France, Institut Pasteur de Lille, évaluation-orientation de la coopération scientifique (ECOS)-Sud [U08S02], and Institut National du Cancer (INCa, projets libres 2008) under reference R08046EE/RPT08003EEA and Agence Nationale de la recherche (ANR) under reference ANR-08-MIEN-021-01 [R08066ES RPV08036ESA]. SI, CP and EMF were recipients of a doctoral fellowship from the Ministère de l'Education Nationale de la Recherche et Technique (SI and EMF) and the Conseil Régional Nord Pas de Calais/Inserm (CP). CF and JCS are supported by Inserm and BR, AB, and FT by the CNRS., Centre National de la Recherche Scientifique (CNRS)-Centre Hospitalier Régional Universitaire [Lille] (CHRU Lille)-Université de Lille-Institut National de la Santé et de la Recherche Médicale (INSERM)-Institut Pasteur de Lille, Réseau International des Instituts Pasteur (RIIP)-Réseau International des Instituts Pasteur (RIIP), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Universidad de la República [Montevideo] (UCUR), Institut des Hautes Etudes Scientifiques (IHES), Centre National de la Recherche Scientifique (CNRS)-Université de Lille, Droit et Santé-Université de Lille, Sciences et Technologies, and Sirard, Jean-Claude

Subjects
MESH: Signal Transduction, Male, MESH: Interleukin-17, Kaplan-Meier Estimate, medicine.disease_cause, Mice, 0302 clinical medicine, Immunology and Allergy, Macrophage, MESH: Animals, MESH: Pneumococcal Infections, MESH: Antigens, CD, 0303 health sciences, education.field_of_study, MESH: Dendritic Cells, Interleukin-17, MESH: Galactosylceramides, 3. Good health, Pneumococcal infections, Infectious Diseases, medicine.anatomical_structure, Streptococcus pneumoniae, [SDV.IMM]Life Sciences [q-bio]/Immunology, MESH: Immunity, Innate, Interleukin 17, Bronchoalveolar Lavage Fluid, Integrin alpha Chains, MESH: Streptococcus pneumoniae, Signal Transduction, [SDV.IMM] Life Sciences [q-bio]/Immunology, MESH: Interferon-gamma, Population, chemical and pharmacologic phenomena, Galactosylceramides, Biology, Pneumococcal Infections, Microbiology, 03 medical and health sciences, Interferon-gamma, Antigen, MESH: Mice, Inbred C57BL, Antigens, CD, medicine, Animals, education, MESH: Mice, MESH: Kaplan-Meier Estimate, 030304 developmental biology, Innate immune system, Lung, MESH: Bronchoalveolar Lavage Fluid, MESH: Integrin alpha Chains, Dendritic Cells, medicine.disease, MESH: Male, Immunity, Innate, Mice, Inbred C57BL, MESH: Natural Killer T-Cells, Immunology, Natural Killer T-Cells, 030215 immunology
Abstract

International audience; BACKGROUND: Exogenous activation of pulmonary invariant natural killer T (iNKT) cells, a population of lipid-reactive αβ T lymphocytes, with use of mucosal α-galactosylceramide (α-GalCer) administration, is a promising approach to control respiratory bacterial infections. We undertook the present study to characterize mechanisms leading to α-GalCer-mediated protection against lethal infection with Streptococcus pneumoniae serotype 1, a major respiratory pathogen in humans. METHODS AND RESULTS: α-GalCer was administered by the intranasal route before infection with S. pneumoniae. We showed that respiratory dendritic cells (DCs), most likely the CD103(+) subset, play a major role in the activation (IFN-γ and IL-17 release) of pulmonary iNKT cells, whereas alveolar and interstitial macrophages are minor players. After challenge, S. pneumoniae was rapidly (4 hours) eliminated in the alveolar spaces, a phenomenon that depended on respiratory DCs and neutrophils, but not macrophages, and on the early production of both IFN-γ and IL-17. Protection was also associated with the synthesis of various interferon-dependent and IL-17-associated genes as revealed by transcriptomic analysis. CONCLUSIONS: These data imply a new function for pulmonary CD103(+) DCs in mucosal activation of iNKT cells and establish a critical role for both IFN-γ and IL-17 signalling pathways in mediating the innate immune response to S. pneumoniae.

Published
2012

42. Gene delivery with polycationic fullerene hexakis-adducts

Author

Marc Nothisen, Jean-Serge Remy, David Sigwalt, Michel Holler, Julien Iehl, Jean-François Nierengarten, Emmanuelle Morin, Conception et application de molécules bioactives (CAMB), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Chimie des Systèmes Fonctionnels, Centre National de la Recherche Scientifique (CNRS), Laboratoire d'innovation moléculaire et applications (LIMA), and Université de Strasbourg (UNISTRA)-Université de Haute-Alsace (UHA) Mulhouse - Colmar (Université de Haute-Alsace (UHA))-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Fullerene, Polymers, Genetic Vectors, Gene delivery, 010402 general chemistry, 01 natural sciences, Catalysis, Adduct, chemistry.chemical_compound, Cations, Materials Chemistry, Organic chemistry, Humans, 010405 organic chemistry, Metals and Alloys, Gene Transfer Techniques, General Chemistry, [SDV.SP]Life Sciences [q-bio]/Pharmaceutical sciences, Combinatorial chemistry, Quantitative Biology::Genomics, 0104 chemical sciences, Surfaces, Coatings and Films, Electronic, Optical and Magnetic Materials, chemistry, Ceramics and Composites, Fullerenes, DNA, HeLa Cells
Abstract

Polyplexes prepared from DNA and globular compact polycationic derivatives constructed around a fullerene hexakis-adduct core have shown remarkable gene delivery capabilities.

Published
2011

43. Hydrophobic forces and hydrogen bonds in the adhesion between retinoid-coated surfaces

Author

Frederic Pincet, Luc Lebeau, Eric Perez, David Tareste, Institut Jacques Monod (IJM (UMR_7592)), Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Physique Statistique de l'ENS (LPS), Université Paris Diderot - Paris 7 (UPD7)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Centre National de la Recherche Scientifique (CNRS)-Fédération de recherche du Département de physique de l'Ecole Normale Supérieure - ENS Paris (FRDPENS), École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS), Conception et application de molécules bioactives (CAMB), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Fédération de recherche du Département de physique de l'Ecole Normale Supérieure - ENS Paris (FRDPENS), École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS), Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Erhart, Marianne, Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Paris (ENS Paris)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Paris (ENS Paris)-Université Pierre et Marie Curie - Paris 6 (UPMC)-Université Paris Diderot - Paris 7 (UPD7)-Centre National de la Recherche Scientifique (CNRS), Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Institut Gilbert-Laustriat : Biomolécules, Biotechnologie, Innovation Thérapeutique, and Université Louis Pasteur - Strasbourg I-Centre National de la Recherche Scientifique (CNRS)

Subjects
Stereochemistry, Surface Properties, Lipid Bilayers, Static Electricity, Dispersive adhesion, 02 engineering and technology, 010402 general chemistry, 01 natural sciences, Hydrophobic effect, Retinoids, Molecular recognition, Cations, Electrochemistry, [CHIM]Chemical Sciences, Non-covalent interactions, General Materials Science, Tromethamine, ComputingMilieux_MISCELLANEOUS, Spectroscopy, [PHYS]Physics [physics], chemistry.chemical_classification, [CHIM.ORGA]Chemical Sciences/Organic chemistry, Chemistry, Chemistry, Physical, Adhesiveness, Water, Hydrogen Bonding, Surfaces and Interfaces, Adhesion, Interbilayer forces in membrane fusion, Hydrogen-Ion Concentration, [CHIM.ORGA] Chemical Sciences/Organic chemistry, 021001 nanoscience & nanotechnology, Condensed Matter Physics, Lipids, 0104 chemical sciences, Folding (chemistry), Quaternary Ammonium Compounds, Crystallography, Chaotropic agent, lipids (amino acids, peptides, and proteins), Salts, Adsorption, 0210 nano-technology
Abstract

Interactions between hydrophobic chains of lipid monolayers and interactions between hydrophilic headgroups of lipid bilayers (with or without a molecular recognition step) are now well documented, especially for commonly used lipids. Here, we report force measurements between a new class of fluorinated lipid layers whose headgroups (synthetic ligands of retinoid receptors) display a very unusual polar/apolar character and can interact via a combination of hydrophobic forces and hydrogen bonds. Although these two interactions produce adhesion and are therefore not easily distinguishable, we show that it is possible to extract both contributions unambiguously. Experiments are performed both in pure water, where the adhesion is a combination of hydrophobic forces and hydrogen bonds, and in Tris buffer, where the hydrophobic effect is the dominant short-range attractive force. The contribution of hydrophobic forces scaled down to molecular interactions is deduced from force versus distance profiles, and the same value is found independently in pure water and Tris buffer, about 1 kBT. We also show that retinoid lipid layers attract each other through a very long-range (100 nm) exponential force, which is insensitive to the pH and the salinity. The origin of this long-range attraction is discussed on the basis of previously proposed mechanisms.

Published
2007

44. Hiyama Cross-Coupling of Chloro-, Fluoro-, and Methoxypyridyltrimethylsilanes: Room-Temperature Novel Access to Functional Bi(het)aryl

Author

Philippe Pierrat, Philippe C. Gros, Yves Fort, Conception et application de molécules bioactives (CAMB), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Structure et Réactivité des Systèmes Moléculaires Complexes (SRSMC), Institut de Chimie du CNRS (INC)-Université de Lorraine (UL)-Centre National de la Recherche Scientifique (CNRS), PIERRAT, Philippe, Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), and McGill University = Université McGill [Montréal, Canada]

Subjects
Trimethylsilyl, 010405 organic chemistry, Stereochemistry, [CHIM.ORGA]Chemical Sciences/Organic chemistry, Aryl, Organic Chemistry, Halide, [CHIM.CATA] Chemical Sciences/Catalysis, General Medicine, [CHIM.CATA]Chemical Sciences/Catalysis, 010402 general chemistry, Ring (chemistry), [CHIM.ORGA] Chemical Sciences/Organic chemistry, 01 natural sciences, Biochemistry, Medicinal chemistry, 0104 chemical sciences, chemistry.chemical_compound, chemistry, Pyridine, Physical and Theoretical Chemistry, ComputingMilieux_MISCELLANEOUS
Abstract

The incorporation of chloro, fluoro, or methoxy substituents on the pyridine ring of pyridyltrimethylsilanes allowed us to perform efficient Hiyama cross-coupling with various (het)aryl halides. The reactions proceeded smoothly at room temperature leading to the corresponding functional bis(het)aryl in fair to excellent yields. The presence of pyridine nitrogen alpha to the trimethylsilyl group was requisite to achieve the cross-coupling. [Reaction: see text]

Published
2005

45. Interaction of the mitotic inhibitor monastrol with human kinesin Eg5

Author

Salvatore DeBonis, Luc Lebeau, Isabelle Crevel, Anne Blangy, Christine Ebel, Jean-Pierre Simorre, David D. Hackney, Pierre Gans, Dimitrios A. Skoufias, Richard H. Wade, Frank Kozielski, Robert A. Cross, Institut de biologie structurale (IBS - UMR 5075 ), Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Direction de Recherche Fondamentale (CEA) (DRF (CEA)), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Centre National de la Recherche Scientifique (CNRS), Marie Curie Research Institute, MRCI, Conception et application de molécules bioactives (CAMB), Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS), Centre de recherches de biochimie macromoléculaire (CRBM), Université Montpellier 1 (UM1)-Université Montpellier 2 - Sciences et Techniques (UM2)-IFR122-Centre National de la Recherche Scientifique (CNRS), Department of Biological Sciences [Pittsburgh], Carnegie Mellon University [Pittsburgh] (CMU), The Beatson Institute for Cancer Research, University of Glasgow, Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019])-Institut de Recherche Interdisciplinaire de Grenoble (IRIG), Commissariat à l'énergie atomique et aux énergies alternatives (CEA)-Commissariat à l'énergie atomique et aux énergies alternatives (CEA), and Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS)

Subjects
[SDV]Life Sciences [q-bio], Dynein, Kinesins, Biology, Sodium Chloride, 01 natural sciences, Biochemistry, Microtubules, 03 medical and health sciences, chemistry.chemical_compound, Inhibitory Concentration 50, Adenosine Triphosphate, Microtubule, [CHIM]Chemical Sciences, Animals, Humans, Microscopy, Interference, ortho-Aminobenzoates, Mitosis, Nuclear Magnetic Resonance, Biomolecular, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology, Sequence Deletion, 0303 health sciences, Microscopy, Video, Dose-Response Relationship, Drug, 010405 organic chemistry, Dyneins, Thiones, Stereoisomerism, Growth Inhibitors, Peptide Fragments, 0104 chemical sciences, Mitotic inhibitor, Spindle apparatus, Protein Structure, Tertiary, Adenosine Diphosphate, Enzyme Activation, Monastrol, Pyrimidines, chemistry, FOS: Biological sciences, Biophysics, Kinesin, Cattle, 69999 Biological Sciences not elsewhere classified, Binding domain, HeLa Cells
Abstract

The microtubule-dependent kinesin-like protein Eg5 from Homo sapiens is involved in the assembly of the mitotic spindle. It shows a three-domain structure with an N-terminal motor domain, a central coiled coil, and a C-terminal tail domain. In vivo HsEg5 is reversibly inhibited by monastrol, a small cell-permeable molecule that causes cells to be arrested in mitosis. Both monomeric and dimeric Eg5 constructs have been examined in order to define the minimal monastrol binding domain on HsEg5. NMR relaxation experiments show that monastrol interacts with all of the Eg5 constructs used in this study. Enzymatic techniques indicate that monastrol partially inhibits Eg5 ATPase activity by binding directly to the motor domain. The binding is noncompetitive with respect to microtubules, indicating that monastrol does not interfere with the formation of the motor-MT complex. The binding is not competitive with respect to ATP. Both enzymology and in vivo assays show that the S enantiomer of monastrol is more active than the R enantiomer and racemic monastrol. Stopped-flow fluorometry indicates that monastrol inhibits ADP release by forming an Eg5-ADP-monastrol ternary complex. Monastrol reversibly inhibits the motility of human Eg5. Monastrol has no inhibitory effect on the following members of the kinesin superfamily: MC5 (Drosophila melanogaster Ncd), HK379 (H. sapiens conventional kinesin), DKH392 (D. melanogaster conventional kinesin), BimC1-428 (Aspergillus nidulans BimC), Klp15 (Caenorhabditis elegans C-terminal motor), or Nkin460GST (Neurospora crassa conventional kinesin).

Published
2003
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46. The interaction between lipid derivatives of colchicine and tubulin: Consequences of the interaction of the alkaloid with lipid membranes

Author

Pierre Ducray, Eric Trudel, Inge Erk, Charles Mioskowski, Luc Lebeau, Marie-France Carlier, Françoise Vérétout, Jean Lepault, Christian Salesse, Stéphane Mons, Laboratoire d'enzymologie et biochimie structurales (LEBS), Centre National de la Recherche Scientifique (CNRS), Virologie moléculaire et structurale (VMS), Institut National de la Recherche Agronomique (INRA)-Centre National de la Recherche Scientifique (CNRS), Institut Gilbert-Laustriat : Biomolécules, Biotechnologie, Innovation Thérapeutique, Université Louis Pasteur - Strasbourg I-Centre National de la Recherche Scientifique (CNRS), Conception et application de molécules bioactives (CAMB), Université de Strasbourg (UNISTRA)-Institut de Chimie du CNRS (INC)-Centre National de la Recherche Scientifique (CNRS), Centre National de la Recherche Scientifique (CNRS)-Université de Strasbourg (UNISTRA), and Institut de Chimie du CNRS (INC)-Université de Strasbourg (UNISTRA)-Centre National de la Recherche Scientifique (CNRS)

Subjects
Photoisomerization, Polymers, Membrane lipids, [SDV]Life Sciences [q-bio], Lipid Bilayers, Biophysics, Biochemistry, Fluorescence, GTP Phosphohydrolases, 03 medical and health sciences, chemistry.chemical_compound, Membrane Lipids, Isomerism, X-Ray Diffraction, In vivo, Tubulin, Lipid membrane, Electron microscopy, Colchicine, [CHIM]Chemical Sciences, Tubulin immobilization, Lipid bilayer, ComputingMilieux_MISCELLANEOUS, 030304 developmental biology, 0303 health sciences, biology, Molecular Structure, Alkaloid, 030302 biochemistry & molecular biology, Colchicine derivative, Cell Biology, Membrane, chemistry, Microscopy, Fluorescence, biology.protein, Solvents, lipids (amino acids, peptides, and proteins), Crystallization
Abstract

Colchicine is a potent antimitotic poison which is well known to prevent microtubule assembly by binding tubulin very tightly. Colchicine also possesses anti-inflammatory properties which are not well understood yet. Here we show that colchicine tightly interacts with lipid layers. The physical and biological properties of three different lipid derivatives of colchicine are investigated parallel to those of membrane lipids in the presence of colchicine. Upon insertion in the fatty alkyl chains, colchicine rigidifies the lipid monolayers in a fluid phase and fluidifies rigid monolayers. Similarly X-ray diffraction data show that lecithin–water phases are destabilized by colchicine. In addition, an unexpectedly drastic enhancement of the photoisomerization rate of colchicine into lumicolchicine in the lipid environment is observed and further supports insertion of the alkaloid in membranes. Finally the interaction of colchicine with lipids makes the drug inaccessible to tubulin. The possible in vivo significance of these results is discussed.

Published
2000

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