A dual functional chromogenic calix[4]arene, which contains both hydroxy azophenols and triazoles for colorimetric sensing and cationic recognition, respectively, has been synthesized. This system was further functionalized with dimethallylsilane to permit grafting to a glass surface. This grafting may be used to produce a surface modified glass slides that act as reusable ion-selective colorimetric sensors. UV/Vis spectral studies revealed that this compound is capable of binding the calcium cation in aqueous media and producing an easily detectable color change. The same is true for the modified surface setup. For the free receptor in acetonitrile solution, the absorption maximum undergoes a red shift from 390 nm to 496 nm upon exposure to calcium cations. In the case of the modified surface, exposure to acetonitrile solution of calcium perchlorate leads to a change in color from orange to red. Washing with acetone serves to remove the metal cations from the calix[4]arene-functionalized glass and restore the original orange color. The system is then ready for reuse. Specifically, following treatment with acetone, the glass may be submerged in a metal solution once again. In the case of exposure to calcium salts, this re-exposure results in a color change analogous to what was seen for a pristine system. In accord with design expectations, this process may be repeated several times. On the basis of this reproducibility, we think this system meets the reusability requirements necessary for the development of a practical sensor device. / text

No description available.

La tomographie par émission de positons (TEP) est une technique d'imagerie médicale largement utilisée en oncologie. Cette technique peut, par exemple, fournir des informations sur la localisation de tumeurs dans le corps humain, et le développement de nouvelles méthodes pour la production automatisée de radiotraceurs spécifiques reste un domaine d’actualité. Dans ce contexte, un nouveau type de précurseurs silylés pouvant être fluorés en dernière étape a été développé. La fluoration de ce nouveau motif imidazole-silylé par du [19F]F- puis par du [18F]F- a été validée sur différents bioconjugués. Le greffage de ce motif a ensuite été testé selon plusieurs méthodes. En particulier, une des voies de synthèse a pu montrer que la fluoration de bioconjugués portant ce motif et greffés sur une résine était possible au [19F]F-. Enfin, le produit radiomarqué au [18F]F- a pu être détecté au cours d’essais préliminaires, permettant ainsi de valider la stratégie choisie. / Positron Emission Tomography (PET) is a medical imaging technique widely used in oncology. For example, this technique can provide information on the localization of growing tumors in the human body, and the development of news methods for the automated production of specific radiotracers is still required. In this context, a new silylated precursor, which can be fluorinated in the last step, has been developed. The fluorination of this new imidazole-silylated unit with [19F]F- and with [18F]F- has been validated on various bioconjugates. The grafting of this moiety has been tested following different strategies. In particular, one of the synthetic pathway showed that the fluorination of bioconjugates with resin grafted moiety was possible with [19F]F-. Finally, the [18F] radiolabeled product was observed during preliminary [18F]F- fluorination experiments, thus validating the chosen strategy.

Two α-amino acids labelled with 18F, 2-[18F]fluoro-L-tyrosine and 6-[18F]fluoro-L-dopa, are routinely produced at the CRC for PET clinical investigation in oncology and neurology. The first one is a tracer for the in vivo quantitative assessment of cerebral protein synthesis (tumor seeking agent) and the second one is a tracer for in vivo cerebral studies of presynaptic dopaminergic functions. Routine production of these two α-amino acids has been conducted in our laboratory, using a nucleophilic, multistep radiosynthesis approach. The no-carrier-added (nca) [18F]fluoride, which is available in large amounts from proton irradiation of 18O-enriched water in a cyclotron, allows the labelling of benzaldehyde derivatives by the substitution of a good leaving group. The synthesis of [18F]fluorobenzyl bromide derivative by reduction and bromination with gaseous HBr followed by the enantioselective alkylation under Phase Transfer Catalysis (PTC) of a benzophenone imine provides the protected radiopharmaceutical which is hydrolysed and purified by HPLC. Due to the fact that gaseous HBr is quite cumbersome to handle and that its complete automation is difficult, the first goal of our work is to evaluate new methods for the synthesis of the [18F]fluorobenzyl bromide derivatives starting from the labelled benzaldehyde derivatives. In order to propose a radiochemical reaction which can be easily automated, we have evaluated different reactions on solid (and non solid) supported reagents. Five reagents were tested. N-piperidinoaminomethylpolystyrene hydrobromide resin gives the the [18F]fluorobenzyl bromide derivatives as unique products in good yields. In the second part of our work, a new approach for the labelling of peptides from the key intermediate p-[18F]fluorobenzyl bromide has been evaluated using the concept of click chemistry. 1,3-dipolar cycloaddition provides fast access to a large number of five-membered heterocycles. We have used this cycloaddition to label protected p-ethynyl-L-phenylalanine with p-[18F]fluorobenzyl azide. This interesting labelling method will be applied to the labelling of a peptide containing this α-amino acid residue.

No description available.

Applications in controlled release and delivery calls for a good understanding of molecular transport within the carrier material and the dominating release mechanisms. It is clear that a better understanding of hindered transport and diffusion of guest molecules is important when developing new porous materials, e.g., surfactant templated silica spheres, for biotechnological applications. Confocal laser scanning microscopy was used to quantify the bulk release and intraparticle transport of small charged fluorescent dyes, and fluorescently-tagged neutral dextran, from mesoporous silica spheres. The time dependent release and the concentration profiles within the spheres have been used to analyze the release mechanisms using appropriate models. While the small, non-adsorbing anionic dye is released following a simple diffusion driven process, the concentration of the cationic dye varies radially within the spheres after loading. The release of the cationic dye is controlled by diffusion after an initial period of rapid release, which could be due to a significant fraction of the cationic dye that remains permanently attached to the negatively charged walls of the mesoporous silica spheres. The diffusion of dextran and the resulting flat concentration profiles could be related to the complex structural feature of the cylindrical pores close to the surface, and a possible conformational change of the dextran with the concentration. The stability and leaching of a catalytic enzyme, lipase, immobilized in hydrophobilized mesoporous support has also been quantified. Colloidal monodisperse mesoporous silica spheres were synthesized and transmission electron microscopy showed that the inner pore structure display a radially extending pores. The mesoporous spheres were used as solid supports for a lipid membrane incorporated with a multi-subunit redox-driven proton pump, which was shown to remain functional. / Synthesis, functionalisation and controlled release of mesoporous materials

Ce manuscrit décrit la synthèse et l'évaluation biologique d'analogues de l'ω-agatoxine IVB dans le but de trouver de nouveaux outils pour l'étude de l'activité des canaux calciques. L'ω-agatoxine IVB est une neurotoxine peptidique isolée du venin d'araignée Agelenopsis aperta qui à ce jour est l'inhibiteur spécifique et sélectif des canaux calciques voltage-dépendants de type P/Q. Ces canaux sont impliqués dans la neurotransmission dépendante du Glutamate dans le système nerveux central. La synthèse de structures peptidiques simplifiées, en comparaison avec celle de la toxine native est décrite. La méthodologie de synthèse de différents analogues cycliques de cette neurotoxine est présentée. Les composés sont synthétisés par synthèse peptidique sur phase solide en stratégie Fmoc, avec une étude particulière sur les conditions de cyclisation et le choix des groupements protecteurs appropriés. Les modifications d es peptides naturels pour obtenir de nouveaux composés biologiquement actifs incluent l'insertion d'aminoacides non naturels et de liaisons pseudopeptidiques. Les analogues synthétisés ont été testés par des méthodes d'électrophysiologie (patch clamp) et d'imagerie calcium ; les activités biologiques des peptides sont corrélées à l'aide d'analyses structurales par RMN et modélisation moléculaire. / This manuscript describes the synthesis and biological valuation of w-agatoxin IVB mimetics with the intention of finding new tools for the study of calcium channels activity. w-Agatoxin IVB is a peptide neurotoxin isolated from the venom of spider Agelenopsis aperta which is a specific and selective inhibitor of P/Q-type voltage-dependent calcium channels. These channels are involved in Glutamate-dependent neurotransmission in the central nervous system. The synthesis of structurally simplified peptides, in comparison with native toxin is described. The methodology of synthesis of different cyclic analogues of this neurotoxin is presented. The compounds were synthesized by solid phase peptide chemistry and Fmoc strategy, with particular consideration for cyclization conditions and an insight into selection of protecting groups. The modifications of the natural peptide to get new biologically active compounds included the insertion of unnatura l amino acids and pseudopeptides bonds. The synthesized analogues were tested by methods of electrophysiology (patch clamp) and calcium imagery; the biological activities of peptides are compared with the aid of structural analyses by RMN and molecular modeling.

Depuis sa découverte, le monoxyde d'azote n'a pas cessé d'intéresser la communauté scientifique. Sa biosynthèse est catalysée par les NO Synthases, qui est une famille de trois isoenzymes. La NOS neuronale et la NOS endothéliale sont constitutives et produisent du NO impliqué dans la neurotransmission et la vasodilatation respectivement. La NOS inductible quant à elle produit du NO impliqué dans la réponse immunitaire innée. Une surproduction de monoxyde d'azote est impliqué dans de nombreuses pathologies comme les maladies neuro-dégénératives ou les maladies chroniques à composantes inflammatoires. L'inhibition de iNOS et nNOS présente donc un grand intérêt thérapeutique. Cependant l'enjeu est de développer des inhibiteurs hautement sélectifs, afin de préserver les fonctions vitales de eNOS.Nous avons choisi de synthétiser des composés constitués d'un analogue de substrat tel que thiocitrulline et S-alkyl-isothiocitrulline auquel est ajouté, par un lien peptidique ou hétérocyclique, un résidu susceptible de se lier dans le canal d'accès du substrat, région moins conservée du site actif pouvant fournir des interactions spécifiques. La synthèse de ces composés a été réalisée sur support solide selon une méthode d'ancrage par la chaîne latérale développée au laboratoire, ou pour certains composés, selon une méthode classique d'ancrage par l'amine alpha, ces deux méthodes offrant un fort potentiel en chimie combinatoire. Les différents composés synthétisés ont été testés sur les trois isoformes recombinantes.Un second travail est brièvement exposé dans ce manuscrit. La synthèse de cyclopeptides basés sur le motif RGD impliqué dans la liaison aux intégrines a été réalisée selon un mode de cyclisation développé au laboratoire faisant intervenir la formation d'un pont guanidine qui sera ensuite diversement substitué. L'étude RMN et l'évaluation de leur activité biologique sur les récepteur opioïdes semble montrer un influence de la substitution du pont sur leur conformation et leur activité. / Since its discovery, Since its discovery, the nitric oxide did not stop interesting the scientific community. Its biosynthesis is catalyzed by NO synthases, a family of three isoenzymes. Neuronal NOS and endothelial NOS are constitutive and produce NO involved in neurotransmission and vasodilation process respectively. Inducible NOS produces NO involved in the innate immune response. An overproduction of nitric oxide is involved in many diseases such as neurodegenerative diseases or chronic diseases with inflammatory components. Thus, its inhibition should be of high therapeutical interest. However, it is necessary to develop highly selective inhibitors to preserve the vital functions of eNOS.We chose to synthesize compounds constituted of one substrate analogue as thiocitrulline or S-alkyl-isothiocitrulline which linked by a peptide bond or heterocycles to a residue able to bind into the substrate access channel, a less conserved region into the active site were specific interactions could be established. The synthesis of these compounds was performed on solid support according to an anchoring method through the side chain developed at the laboratory or, for some compounds, according to a conventional anchoring method through the alpha amine. These approaches are particulary interesting for a combinatory chemistry approach. All compounds were tested on the three recombinant isoforms.A second work is outlined in this manuscript. It consist of synthesizing cyclopeptides based on RGD motif involved in the binding to integrins and enkephalins analogues. The cyclisation method was developed in the lab and involves the formation of a guanidine bridge diversely substituted. NMR studies and biological evaluations on opioid receptor suggests that the diverse bridge substitutions could modulate the conformation and activity of the peptides.

Die im Jahre 1963 von Robert Bruce Merrifield vorgestellte festphasengebundene Peptidsynthese beeinflusste in hohem Maße verschiedene Bereiche der Naturwissenschaften. Doch trotz zahlreicher neuer Entwicklungen hat sich in der das Prinzip der eingesetzten Trägermaterialien nicht grundlegend geändert. Geringfügig quervernetzte Polystyrol-Harze sind immer noch die meist verwendeten Trägermaterialien in der standardisierten Peptidsynthese. In dieser Arbeit wurden superparamagnetische Kern-Schale-Nanopartikel entwickelt und erstmals deren Einsatz als neues Trägermaterial für die Synthese von Peptiden, Peptid-Polymer-Konjugaten und Oligonucleotiden demonstriert. Unter Verwendung einer mikrowellenunterstützten Syntheseroute gelang es zunächst superparamagnetische Magnetitpartikel mit einem Durchmesser von durchschnittlich 6 nm darzustellen. Anschließend wurde ein neu entwickelter mikrowellenunterstützter Stöber-Prozess zur Herstellung von Magnetit-Silica-Kern-Schale-Nanopartikel angewendet, welche im dritten Schritt mit Aminopropyltrimethoxysilan funktionalisiert wurden. Es wurden hochpräzise, monodisperse Kern-Schale-Nanopartikel mit einem Durchmesser von durchschnittlich 69 nm und einem Beladungswert von 0.11 mmol/g erhalten, welche in durchgeführten Stabilitätstests hervorragende Ergebnisse zeigten und als neue Trägermaterialien für festphasengebundenen Synthesen getestet wurden. Die erforderliche Produktaufreinigung erfolgte durch ein externes Magnetfeld, durch welches die Partikel reversibel sedimentierbar sind. Erste Studien der Synthese einer 4-mer-Peptidsequenz zeigten Ausbeuten von über 70% und mit herausragender Reinheit von über 95%. Besonders eindrucksvolle Ergebnisse erzielten die Partikel bei der Synthese von Peptid-Polymer-Konjugaten, bei denen die Ligationsreaktionen mit vorher nicht dokumentierten Umsatzraten verliefen. Außerdem konnte die Anwendbarkeit bei der Synthese eines Trinucleotids nachgewiesen werden. / In 1963 Merrifield introduced the method of solid-phase supported synthesis and thus revolutionized peptide synthesis. In spite of several new developments, the main principle of established solid supports has not changed much. Still lightly cross linked poly(styrene) resins dominate the used supports. This work reports on surface amino functionalized, superparamagnetic nanoparticles with a protective silica shell to be applicable as colloidal supports for organic synthesis of peptides, peptide polymer conjugates and oligonucleotides. A microwave supported synthesis route lead to superparamagnetic magnetite particles with an average particle diameter of 6 nm. Subsequently a new developed microwave assisted Stöber process was used to build up magnetite-silica-core-shell-nanoparticles, which were functionalized in a third step with aminopropyltrimethoxysilane. Defined monodisperse core-shell nanoparticles were obtained with an average diameter of 69 nm and a concentration of free amino groups of 0.11 mmol/g, which showed excellent results in conducted stability tests and were used as new support materials for solid-supported syntheses. Convenient magnetic sedimentation proved to ensure ease of purification after each reaction step. Initial studies of a synthesis of a tetramer peptide sequence showed yields of more than 70% and an outstanding purity of more than 95%. The particles also showed impressive results in the synthesis of peptide-polymer conjugates, in which the ligation reactions proceeded conversion rates, which had not been published before. In addition, the applicability of the particles was demonstrated in the synthesis of a trinucleotide.

Le squelette 1,3,5-triazépane-2,6-dione est un hétérocycle à sept chainons dérivé de dipeptides et accessible en quatre étapes en solution. Une voie de synthèse en parallèle sur support solide de cet hétérocycle a été élaborée. Cette synthèse, qui repose sur les principes de "catch and release" et de cyclo-clivage, a permis la création d’une chimiothèque de plus d’une centaine de composés. Pour augmenter la diversité du squelette 1,3,5-triazépane-2,6-dione, des modifications post-cyclisation peuvent avoir lieu telles que des réactions de N-mono-alkylation ou de N,N-di-alkylation de l’urée, des réactions d’acylation ou bien des réactions de thionation des fonctions carbonyles. De même, la synthèse des cycles analogues aux 1,3,5-triazépane-2,6-diones des tailles plus importantes a été examinée conduisant à l’obtention de plusieurs macrocycles.Les 1,3,5-triazépane-2,6-diones présentent un fort potentiel pour la recherche de molécules d’intérêt thérapeutique puisque le cycle est rigide, non-planaire et possède une bonne capacité de distribution des pharmacophores dans l’espace. Des molécules de cette famille présentent une activité inhibitrice modérée mais spécifique sur les phospholipases A2 secrétées humaines de type V et X. La recherche de nouveaux inhibiteurs de sPLA2 par une étude de relation structure/activité, par création d’une pince à calcium ou par simulation moléculaire a conduit à l’identification de nouveaux composés actifs. / The 1,3,5-triazepane-2,6-dione scaffold is a seven membered heterocycle derived from dipeptides and accessible in a four steps synthesis in solution. A parallel solid phase synthesis of this heterocycle was developed. This strategy, based on "catch and release" and cyclo-cleavage processes, had created a library containing more than one hundred compounds. To increase the diversity of 1,3,5-triazepane-2,6-dione moieties, some post-cyclisation modifications were performed, e.g. urea N-mono-alkylation or N,N-di-alkylation, acylation, and carbonyl thionation. Synthesis of larger cycles was also investigated and several macrocycles were obtained.The 1,3,5-triazepane-2,6-diones have a strong pharmacological interest, because their cycle is rigid, non-planar and can allow multiple presentation of pharmacophores in space. Some 1,3,5-triazepane-2,6-diones have shown a small but specific activity on the groups V and X of the human secreted phospholipases A2. Structure/activity relationships, clamp synthesis to bind calcium or virtual screening were the strategies used to identify new active compounds.