Made available in DSpace on 2014-08-20T13:42:06Z (GMT). No. of bitstreams: 1 Dissertacao_Gabrie_Olle_ Dalmazo.pdf: 526235 bytes, checksum: 5e78165179f8f9e3164e0269bd617ab9 (MD5) Previous issue date: 2011-02-28 / The tropane alkaloids (TA) pathway has a branch-point controlled by the enzymes tropinone reductase 1 and 2 (TR1 and TR2). Tropinone is the common substrate for these enzymes and is reduced either by TR1 to form tropine, hyoscyamine and scopolamine or by TR2 to form pseudotropine and calystegines. Hyoscyamine and scopolamine are largely used in medicine as anticholinergic, antiemetic, parasympatholytic and anaesthetic. Calystegines mimic different sugars and are potent and specific inhibitors of glucosidases. The function of hyoscyamine, scopolamine and calystegines in the plants is not fully understood. Recent studies suggest that they are involved in the plant defense against pathogens. Regulation of TA biosynthesis in planta has attracted interest not only in view of its applications in the pharmaceutical industry, but also in respect to human nutrition and plant physiology. In the present study a virus-based transgenic approach devised to induce PTGS of tr1 and tr2 in whole transformed Hyoscyamus muticus plants is evaluated. It was observed that a significant reduction in transcript accumulation for tr2 caused a tremendous increase in transcript accumulation for tr1. / Um ponto de bifurcação na rota metabólica de tropano alcalóides (TA) é controlado pelas enzimas tropinona redutase 1 e 2 (TR1 e TR2). Tropinona, o substrato comum para estas enzimas, é reduzido por TR1 para formar tropina, hiosciamina e escopolamina ou por TR2 para formar pseudotropine e calisteginas. Hiosciamina e escopolamina são largamente utilizadas em medicina como anticolinérgicos, antieméticos, parassimpatolíticos e anestésicos. Calisteginas têm estrutura molecular semelhante a açúcares e são potentes inibidores específicos de glucosidases. A função dos alcalóides hiosciamina, escopolamina e calisteginas em plantas não é completamente entendida. Estudos recentes sugerem o envolvimento destes alcalóides na defesa da planta contra patógenos. A regulação da biossíntese de TA na planta tem atraído interesse não apenas quanto à aplicação na indústria farmacêutica, mas também com respeito à nutrição humana e fisiologia de plantas. No presente estudo avaliou-se uma estratégia transgênica, baseada em vírus, para induzir o silenciamento gênico pós-transcricional de tr1 e tr2 em plantas de Hyoscyamus muticus L. Observou-se que reduções significativas no acúmulo de transcritos para tr2 causaram um tremendo aumento no acúmulo de transcritos para tr1.

Cette thèse a porté sur la synthèse de cycloheptatriènes (CHT) silylés et leur désymétrisation pour l’obtention de mimes de sucres. Nous avons ainsi développé une approche synthétique d’analogues de calystégines et mis en évidence un réarrangement inattendu conduisant à un squelette nortropane original. Dans un second temps, nous avons étudié l’équilibre cycloheptatriène-norcaradiène (CHT-NCD) et mis au point une méthodologie de cycloaddition de CHT silylés avec des dérivés nitroso, efficace et hautement stéréosélective, pour la synthèse d’aminocarbasucres via une cascade : électrocyclisation (CHT-NCD)-cycloaddition-ouverture cationique-cycloaddition. En 3 étapes, à partir du CHT méthylsilylé, 7 centres stéréogènes, 5 liaisons C-O et 2 C-N sont formés de façon diastéréo- et régiocontrôlée. Une réaction Hétéro- Diels-Alder énantiosélective sur les CHT silylés, qui conduit en une étape à la formation de 3 centres asymétriques, a aussi été développée. / This work dealt with the synthesis of silylated cycloheptatrienes and their desymmetrisation toward the synthesis of sugar mimics. Calystegines analogues were synthesized and an unexpected rearrangement leading to an original nortropane skeleton was emphasized. In a second part, cycloheptatriene-norcaradiene (CHT-NCD) equilibrium was studied. Silylated cycloheptatriene were also shown to react through cycloaddition reaction with acylnitroso compounds, through cascade processes: electrocyclisation (CHT-NCD)-cycloaddition- cationique cyclopropane opening-cycloaddition, leading highly selectively to aminocarbasugars. Starting from methylsilylated CHT, 7 stereogenic centers, 5 C-O bonds and 2 C-N bonds were formed in only 3 steps. An enantioselective Hetero-Diels-Alder reaction was also developed.