Un très grand nombre de travaux de recherche fait état de l’intérêt des oligonucléotides en tant qu’agents thérapeutiques. Les modes d’actions envisageables sont très variés (thérapie antisens, antigène, interférence ARN, etc.). Cependant, les propriétés pharmacocinétiques et pharmacodynamiques des oligonucléotides naturels ne permettent pas leurs utilisations in vivo. Leurs propriétés peuvent être améliorées par des modifications chimiques. Notre travail consiste à synthétiser une nouvelle génération d’oligoribonucléotides modifiés : les oligomères de nucléosides aminooxy acides. Dans ces oligomères, la liaison phosphodiester de l’ARN est remplacée par une liaison N-oxyamide -CONHO-. Cette liaison est stable vis-à-vis des hydrolyses chimiques et enzymatiques et est facilement engagée dans des liaisons hydrogène. La préparation de différents nucléosides aminooxy esters protégés à partir de l’uridine ou du D-(+)-glucose est présentée. Par ailleurs, les N-oxy PNA constituent une autre famille d’oligonucléotides modifiés présentant une liaison N-oxyamide. L’analyse structurale des monomères et des dimères de N-oxy PNA est détaillée.De plus, un projet en collaboration avec le LBPA s’intéresse à une méthode de détermination de la structure secondaire des ARN. Dans ce but, nous avons conçu, synthétisé et étudié des sondes chimiques ciblant le ribose des nucléotides non appariés d’ARN. L’emploi de catalyseurs nucléophiles comme la DMAP permet d’augmenter la réactivité des sondes. / A large number of researches report the interest of oligonucleotides as therapeutic agents. The modes of actions are very varied (antisense therapy, antigen therapy, RNA interference, etc.). However, the pharmacokinetic and pharmacodynamic properties of natural oligonucleotides do not allow their in vivo uses. Their properties can be improved by chemical modifications. Our work consists to synthesize a new generation of modified oligoribonucleotides: the oligomers of aminooxy acids nucleosides. In such oligomers, the phosphodiester bond of the RNA is replaced with a N-oxyamide bond -CONHO-. This linkage is stable to chemical and enzymatic hydrolysis and is easily engaged in hydrogen bondings. The preparation of different protected aminooxy esters nucleosides starting from uridine or D-(+)-glucose is presented. Furthermore, N-oxy PNA constitute another family of modified oligonucleotides having a N-oxyamide bond. Structural analysis of the monomers and the dimers of N-oxy PNA is detailed.In addition, a project in collaboration with the LBPA focuses on a method for determining the secondary structure of RNA. To this end, we designed, synthesized and studied chemical probes targeting ribose of unpaired nucleotides. The use of nucleophilic catalysts such as DMAP increases the reactivity of the probes.

Les ARN interférents sont de puissants outils thérapeutiques et biologiques pour la mise en silence de l'expression des gènes. Afin d'améliorer leur stabilité enzymatique, leur biodistribution et leur pénétration cellulaire, nous proposons de développer une approche prodrogue d'ARN interférent. Ce manuscrit rapporte la synthèse et l'évaluation de pro-ARN masqués temporairement par des groupements biolabiles susceptibles d'être hydrolysés dans les cellules afin de libérer l'ARN naturel actif. Deux types de modifications sont présentés : des groupes acétalesters enlevés par des carboxyestérases et des groupes alkyldithiométhyles sensibles à un environnement réducteur. Dans une première partie, une nouvelle méthode de synthèse de pro-ARN partiellement modifiés en position 2' par des groupements acétalesters est décrite. Plusieurs groupements variant par leur caractère lipophile ou cationique sont évalués. Des résultats prometteurs d'études physico-chimiques, de stabilité enzymatique, de pénétration cellulaire et d'inhibition de gènes mettent en valeur l'intérêt d'utiliser certains pro-ARN modifiés en tant qu'outils thérapeutiques. Une deuxième partie présente une voie de synthèse originale de pro-ARN modifiés en position 2' par des groupements alkyldithiométyles. Les propriétés physico-chimiques, la stabilité enzymatique et le démasquage de ces pro-ARN sont décrits. Parallèlement, l'étude d'une réaction d'échange thiol-disulfure permettant l'incorporation de liens disulfures intrabrin au sein de duplex d'ARN et de constructions tige-boucles est détaillée dans ce manuscrit. / SiRNA are powerful therapeutic and biological tools for gene silencing. In the aim of improving their stability, their biodistribution and their cellular delivery, we propose to develop a siRNA prodrug-like approach.This manuscript reports the synthesis and the study of pro-RNA temporarily masked by biolabile groups which could be hydrolyzed inside cells in order to release the active unmodified RNA. Two types of modifications are presented: acetalester groups removed by carboxyesterases and alkyldithiomethyl groups cleaved in a reducing environment within cells.In a first part, a new synthesis strategy of partially modified 2'-O-acetalester pro-RNA is described. Several acetalester groups varying in their lipophilicity and their charge are evaluated. Promising results obtained in physical-chemical studies, enzymatic stability and gene inhibition highlight the use of these modified pro-RNA as therapeutic drugs. A second part introduces an original approach for the synthesis of 2'-O-alkyldithiomethyl pro-RNA. The physical-chemical properties, the enzymatic stability and the unmasking of this pro-RNA are described.Moreover, the study of a thiol-disulfide exchange reaction allowing the incorporation of intrastrand disulfide bond into secondary structure duplex and hairpin is reported in this manuscript.

Compostos carcinogênicos estruturalmente diversos ligam-se covalentemente ao DNA formando adutos que, se não reparados, provocam mutações. Inicialmente relacionados apenas a compostos exógenos, atualmente há várias evidências de que compostos gerados endogenamente poderiam modificar o DNA gerando adutos. Dentre os compostos endógenos, os produtos carbonílicos α,β-insaturados destacam-se pois reagem como agentes alquilantes bifuncionais com as bases do DNA, formando adutos exocíclicos. O 2,4-decadienal (DDE) é um aldeído α,β-insaturado que além de estar presente em alimentos e poluentes, é um dos mais importantes produtos da lipoperoxidação. Embora há várias indicações sobre a ação genótoxica do DDE, nenhum aduto deste composto com nucleobases havia sido caracterizado. O presente trabalho mostrou que o DDE é um agente alquilante versátil sendo capaz de gerar cinco adutos diferentes. Este estudo também mostrou que o DDE é capaz de gerar os mesmos adutos que outros dois importantes produtos da lipoperoxidação: o 4-0H-nonenal e o 2-octena1. Todos os adutos foram detectados em DNA tratado in vitro com o DDE. Para possibilitar a detecção dos adutos em sistemas mais complexos, foi desenvolvido um método extremamente sensível baseado em HPLC interfaceado com espectrometria de massa em tandem com ionização por electrospray. Foi também desenvolvida uma estratégia de incorporação de um dos adutos (III) em oligonucleotídeos por via química. A estratégia foi utilizada com sucesso na incorporação do aduto em oligonucleotídeos de sequências diversas. Os oligonucleotídeos foram utilizados em ensaios de reparo por excisão de base e replicação in vitro. / Structurally diverse carcinogenic compounds bind covalently to DNA producing adducts that can, if not repaired, lead to mutations. Firstly restricted to exogenous compounds, nowadays there are evidences that compounds endogenously generated can modify DNA forming adducts. Among these endogenous compounds, α,β-unsaturated carbonyl products are of special interest due their reactivity as bifunctional alkylating agents towards DNA bases leading to exocyclic adducts. 2,4-decadienal (DDE) is an α,β-unsaturatedaldehyde that in addition to be present in food and pollutants, it is one of the most important lipid peroxidation products. Despite of many indications about the DDE genotoxic properties, there was no information about adducts produced between this compounds and nucleobases. This work showed DDE as a versatile DNA alkylating agent being able to generate five different adducts. This study also showed that DDE can generate the same adducts produced from two other important lipid peroxidation products: 4-0H-nonenal and 2-octena1. All adducts were detected in DNA treated in vitro with DDE. To allow adduct detection in more complex systems, we developed a highly sensitive method based on HPLC coupled to electrospray/tandem mass spectrometry. A strategy to incorporate one adduct (III) into oligonucleotides was also developed. The strategy was successfully applied in the adduct incorporation into diverse sequences of oligonucleotides. They were utilized in base excision repair and in vitro replication experiments.

Compostos carcinogênicos estruturalmente diversos ligam-se covalentemente ao DNA formando adutos que, se não reparados, provocam mutações. Inicialmente relacionados apenas a compostos exógenos, atualmente há várias evidências de que compostos gerados endogenamente poderiam modificar o DNA gerando adutos. Dentre os compostos endógenos, os produtos carbonílicos α,β-insaturados destacam-se pois reagem como agentes alquilantes bifuncionais com as bases do DNA, formando adutos exocíclicos. O 2,4-decadienal (DDE) é um aldeído α,β-insaturado que além de estar presente em alimentos e poluentes, é um dos mais importantes produtos da lipoperoxidação. Embora há várias indicações sobre a ação genótoxica do DDE, nenhum aduto deste composto com nucleobases havia sido caracterizado. O presente trabalho mostrou que o DDE é um agente alquilante versátil sendo capaz de gerar cinco adutos diferentes. Este estudo também mostrou que o DDE é capaz de gerar os mesmos adutos que outros dois importantes produtos da lipoperoxidação: o 4-0H-nonenal e o 2-octena1. Todos os adutos foram detectados em DNA tratado in vitro com o DDE. Para possibilitar a detecção dos adutos em sistemas mais complexos, foi desenvolvido um método extremamente sensível baseado em HPLC interfaceado com espectrometria de massa em tandem com ionização por electrospray. Foi também desenvolvida uma estratégia de incorporação de um dos adutos (III) em oligonucleotídeos por via química. A estratégia foi utilizada com sucesso na incorporação do aduto em oligonucleotídeos de sequências diversas. Os oligonucleotídeos foram utilizados em ensaios de reparo por excisão de base e replicação in vitro. / Structurally diverse carcinogenic compounds bind covalently to DNA producing adducts that can, if not repaired, lead to mutations. Firstly restricted to exogenous compounds, nowadays there are evidences that compounds endogenously generated can modify DNA forming adducts. Among these endogenous compounds, α,β-unsaturated carbonyl products are of special interest due their reactivity as bifunctional alkylating agents towards DNA bases leading to exocyclic adducts. 2,4-decadienal (DDE) is an α,β-unsaturatedaldehyde that in addition to be present in food and pollutants, it is one of the most important lipid peroxidation products. Despite of many indications about the DDE genotoxic properties, there was no information about adducts produced between this compounds and nucleobases. This work showed DDE as a versatile DNA alkylating agent being able to generate five different adducts. This study also showed that DDE can generate the same adducts produced from two other important lipid peroxidation products: 4-0H-nonenal and 2-octena1. All adducts were detected in DNA treated in vitro with DDE. To allow adduct detection in more complex systems, we developed a highly sensitive method based on HPLC coupled to electrospray/tandem mass spectrometry. A strategy to incorporate one adduct (III) into oligonucleotides was also developed. The strategy was successfully applied in the adduct incorporation into diverse sequences of oligonucleotides. They were utilized in base excision repair and in vitro replication experiments.

In this study, the effects of certain chromatographic conditions on various modified and unmodified oligonucleotides were investigated. At the forefront of this study was the investigation of a new Ion-pair Reversed-phase liquid chromatography (IP-RPLC) method, that had the potential to replace a previously established triethylammonium acetate (TEAA) IP-RPLC method developed for oligonucleotide separations. This method, utilising the counter ion dibutyl amine (DBA) and a Tris-buffer at pH 8, produced promising results indicating that the strong binding strength of DBA creates a hybrid IEX/RPLC separation method – the separation of oligonucleotides is dynamically based on both charge and length. Higher concentrations of DBA appear to produce better results that include improved efficiency, increased retention and even the potential discovery of hidden impurities. In conjugation with Ultra-high-pressure liquid chromatography (UHPLC) systems, sub-2 µm particle columns and gradient optimisations, separations of complex oligonucleotides could be achieved in short analysis times. Furthermore, effective separations at the analytical level can be applied and adapted to larger scale Prep-LC, potentially also improving the purification process of crude oligonucleotide samples. Further development and validation are, however, required for any future work with this method. / I denna studie har effekten av vissa kromatografiska förhållanden på olika modifierade och icke-modifierade oligonukleotider undersökts. I framkanten av denna studie var undersökningen av en ny IP-RPLC metod, vilken har potential att ersätta den tidigare etablerade trietylammonium acetat (TEAA) IP-RPLC metoden, vilken utvecklats för separationen av oligonukleotider. Denna metod, vilken använder dibutylamin (DBA) som motjon och en Tris-buffert vid pH 8, gav lovande resultat vilka indikerar att den starka bindningsstyrkan av DBA skapar en hybrid IEX/RPLC separationsmetod – separationen av oligonkuleotider styrs både av dess laddning och dess längd. Höga koncentrationer av DBA verkade ge bättre resultat som inkluderar hög effektivitet, ökad retention och även den potentiella upptäckten av gömda föroreningar. I samband med UHPLC systemer, kolonner med mindre än 2µm i partikelstorlek och optimiserade gradienter, separationer av komplexa oligonukleotider erhölls på korta analystider. Effektiva separationer vid den analytiska nivån kan appliceras och adapteras till storskalig preparative-LC, med potential att kunna förbättra reningsprocessen för syntetiserade oligonukleotider. Vidare utveckling och validering krävs för framtida användning av denna metod.

Die Telomere sind spezialisierte DNA-Protein-Komplexe, die sich an den Enden der Chromosomen der eukaryotischen Zellen befinden. Die Telomerase ist ein Ribonukleoprotein, welches für die vollständige Replikation der Telomere bei den meisten Eukaryoten verantwortlich ist. Die katalytische Untereinheit des Enzyms (hTERT beim Menschen) besitzt Reverse-Transkriptase-Aktivität, und nutzt eine integrierte RNA (hTR beim Menschen) als Template, um Telomer-Wiederholungssequenzen an den Enden der Chromosomen zu synthetisieren. Die Telomerase ist in den meisten normalen humanen somatischen Zellen unterdrückt. In den meisten Krebszellen jedoch, stellt die Reaktivierung der Telomerase zur Beibehaltung der Telomerlänge eine Voraussetzung für deren unbegrenztes Wachstumspotential dar. Im Rahmen dieser Arbeit sollte ein neuer, einfacher und selektiver Assay für den Nachweis der humanen Telomerase entwickelt werden. In dem neuen Assay sollten die beiden Kernkomponenten des Enzyms, die Protein-Untereinheit und die RNA, die Targets sein. Der Test ist in seiner Grundstruktur wie folgt aufgebaut : 1. Immobilisierung der Telomerase über die hTERT an eine Festphase, beschichtet mit Phosphorothioat-modifizierten (PS) Oligonukleotiden oder Heparin. Zusammen mit der Telomerase werden bei diesem Schritt die Heparin-bindenden Proteine, die in der Probe enthalten sind, an die Festphase gebunden. 2. Spezifischer Nachweis der hTR. Zur Detektion der hTR wird ein Oligonukleotid-Ligations-Assay (OLA) oder eine Reverse-Transkriptase-PCR (RT-PCR) eingesetzt. In der optimierten Endversion wurde zur Immobilisierung des Enzyms eine Festphase, beschichtet mit PS-Oligonukleotiden, verwendet. Die hTR wurde mittels RT-PCR nachgewiesen. Mit dem neuen Assay wurden erfolgreich 75 Tumorzellen detektiert. / Telomeres are specialized DNA-Protein structures located at the ends of linear eukaryotic chromosomes. Telomerase is a ribonucleoprotein, which is responsible for the complete replication of the telomeres in most eukaryotes. The catalytic reverse transcriptase protein subunit (hTERT in humans) of the nucleoprotein uses an integral RNA (hTR in humans) as a template for the addition of telomeric repeat sequences to the ends of chromosomes. Telomerase is repressed in most normal human somatic cells, while the reactivation of telomerase to maintain telomere length is necessary for the unlimited growth potential of most human cancer cells. The aim of this work was the development of a new, simple and selective assay for the detection of human telomerase. The targets of the new assay were the two core subunits of the enzyme : hTERT and hTR. The test comprises two principal steps : 1. Immobilization of the telomerase via the hTERT subunit on a solid phase, coated with heparin or phosphorothioate-modified (PS) oligonucleotides. In this step telomerase is bound together with the heparin-binding proteins of the analysed sample to the surface. 2. Specific detection of the hTR. For the detection of the hTR an oligonucleotide ligation assay (OLA) or a reverse transcriptase PCR (RT-PCR) was used. In the optimized final version of the assay a PS-coated solid phase was used for the immobilization of the enzyme. Reverse transcriptase PCR was applied for detection of the hTR. 75 tumor cells were successfully detected with the new assay.

This thesis is concerned with synthesis, structure and biochemical analysis of chemically modified oligonucleotides with potential therapeutic applications. The three types of chemical modifications described here are: (a) A North-East locked 1',2'-azetidine nucleoside (b) A North locked 2',4'-cyanomethylene bridged nucleoside and (c) A 2',4'-aza-ENA-T nucleoside. The synthesis of the 1',2'-azetidine fused nucleosides was described using two different approaches. A highly strained 2',4'-cyanomethylene locked nucleoside was synthesized but could not be converted to the phosphoramidite derivative due to instability during derivatization. The key cyclization step in the aza-ENA-T nucleoside synthesis gave rise to two separable diastereomers due to chirality at the exocyclic nitrogen. Conversion of diastereomer 55 to 56 occurred with a large free energy of activation (ΔG‡ = 23.4 kcal mol-1 at 298 K in pyridine-d5). Of the two isomers the equatorial NH product was more stable than the axial one due to reduced 1,3 diaxial interactions. As a result, all NH axial product was converted to the equatorial isomer during subsequent steps in the synthesis. NMR and ab initio experiments confirmed the North-East structure of the 1',2'-azetidine locked nucleoside and North conformation of aza-ENA-T locked nucleosides with a chair conformation of the piperidine ring. The amino modified nucleosides were incorporated into different positions of a 15mer oligonucleotide. The azetidine modified AONs did not form stable duplexes with complementary RNA (ΔTm ~-1 to -4 °C), but they performed better than previously synthesized isosequential 1',2'-oxetane modified oligonucleotides. The 2',4'-aza-ENA-T modified oligonucleotide, on the other hand, showed excellent target affinity with complementary RNA (ΔTm ~+4 °C). The azetidine and aza-ENA-T modified oligonucleotides showed significant stability in the presence of human serum and snake venom phosphodiesterase (3'-exonuclease) as compared to the unmodified native sequence. The singly modified 15mer oligonucleotides were also subjected to RNase H promoted digestion in order to evaluate their potential as effective antisense agents. The effective enzyme activity (kcat/Km) was found to be lower in the modified AONs due to reduced enzyme-substrate binding. However, the catalytic activity of RNase H with these modified-AON:RNA duplexes were higher than observed with the native duplex.

A significant focus of toxicological research is the identification of electrophiles that covalently modify DNA to form addition products (adducts). These products can be generated when aryl radical species react at the C8-site of 2'-deoxyguanosine (dG) to form C8-aryl-dG adducts, which are mutagenic lesions. While this form of DNA modification is detrimental, C8-aryl-dG adducts also possess intriguing properties that can be exploited for beneficial purposes. This thesis is an investigation of one mechanism believed to contribute to the mutagenicity of C8-aryl-dG adducts, as well as a study of the photophysical properties of adducts that allow for their application as fluorescent probes. A common property of C8-aryl-dG adduction is accompaniment of abasic site formation. To determine how the C8-aryl moiety contributes to sugar loss, UV-Vis spectroscopy has been employed to determine hydrolysis kinetics, with C8-aryl-dG adducts found to be more prone than dG to acid-catalyzed hydrolysis. Despite adduct reactivity in acidic media, all adducts are relatively stable at pH 7, suggesting they are unlikely intermediates of abasic site formation at physiological pH. These results have allowed for development of a new rationale for depurination observed upon C8-aryl-dG adduction within duplex DNA. The determination of photophysical parameters of C8-heteroaryl-dG adducts reveals that these nucleosides behave as fluorophores with high fluorescence quantum yields (φfl). These adducts also exhibit emission sensitivity to their solvent environment and H-bonding interactions. C8-Heteroaryl-dG adducts were incorporated in the oligonucleotide 5'-CTCG1G2CG3CCATC, at the G1 and G3 sites, that contains the recognition sequence of the NarI Type II restriction enzyme. Hybridization of the modified NarI oligonucleotides to the complementary strand containing either the C or G nucleobase opposite the adduct allowed for characterization of duplex structures by circular dichroism (CD), UV melting temperature analysis and fluorescence spectroscopy. Results suggest that the C8-heteroaryl-dG adduct favours an anti conformation with base-paired with C, while a syn conformation is favoured when base-paired to G. Adduct conformation of bulky C8-dG adducts is believed to be correlated with their known mutagenic activity. C8-Heteroaryl-dG modified nucleosides could therefore be used as fluorescent models of these adducts to aid in elucidation of adduct-induced mutagenesis in biological systems. / NSERC

Les défauts de génome mitochondrial provoquent des maladies neuromusculaires, pour lequel aucun traitement efficace n'a été mis au point. La plupart des mutations mitochondriales sont hétéroplasmique, ce qui signifie que l'ADN mitochondrial (ADNmt) de type sauvage et muté coexistent dans la même cellule, et le changement de proportion entre deux types d'ADNmt pourrait rétablir les fonctions mitochondriales. Le but du projet était le développement du système pour cibler l'ARN thérapeutique dans les cellules humaines vivantes. Au cours de ma thèse j'ai synthétisé une série de nouveaux ARN anti-réplicatifs contenant modifications chimiques pour augmenter leur stabilité dans la cellule, et mis au point la nouvelle méthode de synthèse chimique des molécules d'ARN contenant cholestérol fixé par l'intermédiaire d'un pont biodégradable. Ces ARN étaient capable de pénétrer dans les cellules humains, d'être adressées dans les mitochondries et de diminuer la proportion d' ADNmt muté. / Defects in mitochondrial genome cause neuromuscular diseases, for which no efficient therapy has been developed. Since most mitochondrial mutations are heteroplasmic, wild type and mutated mitochondrial DNA (mtDNA) coexist in the same cell, and the shift in proportion between two mtDNA types could restore mitochondrial functions. The aim of the project was development of carrier-free system for targeting the therapeutic mitochondrially importable RNA into living human cells. During my PhD study, I have synthesized a set of new anti-replicative RNAs containing various chemical modifications, aiming to increase their stability in the cell, and developed a new method for the chemical synthesis of RNA molecules containing cholesterol attached through a biodegradable bridge. Cholesterol containing antireplicative RNAs were characterised by efficient cellular uptake, partial colocalisation with mitochondria and ability to decrease the proportion of mutant mtDNA.

Récemment, de nombreux efforts ont été consacrés au développement d’oligonucléotides synthétiques pour des applications thérapeutiques et de diagnostique variées. Les oligonucléotides modifiés peuvent inhiber sélectivement l’expression des gènes en se liant spécifiquement à des séquences d’ADN et/ou d’ARN ciblées à travers les stratégies antigène, antisens ou d’ARN interférent. Les aminoxy peptides forment facilement des structures secondaires bien définies comme des alpha-, béta-, gamma-turns ou des hélices, ce qui nous a inspiré pour concevoir de nouveaux oligonucléotides modifiés dans le but d’étudier leurs propriétés physico-chimiques et biologiques. Au cours de ce travail, la synthèse de nucléosides aminoxy acides et de leurs oligomères a été entreprise en séries ribose et désoxyribose. Dans la première partie, les fonctions aminoxyle, acide carboxylique et aldéhyde ont été introduites sur la partie osidique de la thymidine. Différents nucléosides monofonctionnalisés ont été synthétisés à l’aide notamment des réactions de Mitsunobu, d’O-allylation et d’oxydation. Les nucléosides monomères ont ensuite été couplés entre eux conduisant aux nouveaux dinucléosides liés par liaison N-oxy amide, oxime et aminoxy. Dans la seconde partie, la synthèse de différentes uridines aminoxy acides a été étudiée à partir de l’uridine, des 2,2’-anhydro et 2,3’-anhydro uridines. Une uridine aminoxy ester a pu être obtenue en passant par la 3’-oxo uridine via une homologation (réaction de Wittig) et l’introduction de la fonction oxyamine en position 5’ par une substitution nucléophile du dérivé iodé. En parallèle, dans la continuité des travaux réalisés au laboratoire sur la synthèse des glycoamino acides, nous avons synthétisé des C-glycosyl aminoxy acides jamais décrits dans la littérature, dans le but de générer de nouveaux mimes de glycopeptides. A partir du C-allyl glucopyranoside perbenzylé, deux C-glucosyl aminoxy acides diastéréoisomères ont été préparés. / Much recent efforts have been devoted to the development of synthetic oligonucleotides for various therapeutic and diagnostic applications because of their capability to cause selective inhibition of gene expression by bonding to the target DNA/RNA sequences through antigen, antisense and RNA interference. The easy formation of well-defined structures like alpha-, béta-, gamma-turns or helical structures of N-oxy peptides promoted us to design new modified oligonucleotides so as to study their physico-chemical and biological properties. During this work, synthesis of different nucleosides aminoxy acids as well as their oligomers has been investigated. In the first part, aminoxy, carboxylic acid and aldehyde functions were introduced into the sugar ring of thymidine and different monofunctionalized nucleosides were obtained thanks to Mitsunobu, O-allylation and oxidation reactions. Different nucleoside monomers were then linked together, leading to novel dinucleosides with N-oxy amide, oxime or aminoxy linkage. In the second part, preparation of different uridines aminoxy acids was studied by using uridine, 2,2’-anhydro and 2,3’-anhydro uridines as starting materials. An uridine aminoxy ester was obtained from 3’-oxo uridine, through homologation with Wittig reaction and introduction of the oxyamine function by nucleophilic substitution of the 5’-iodo derivative. In parallel, as a continuing program in the laboratory on the glycoamino acids synthesis, we have synthesized novel C-glycosyl aminoxy acids in order to generate new mimes of glycopeptides. From perbenzylated C-allyl glucopyranoside, two diastereomeric C-glucosyl aminoxy acids have been successfully prepared.