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Chemoenzymatic Synthesis of Sugar-Related Polyhydroxylated Compounds, Iminocyclitols and Their Derivatives As Glycosidase Inhibitors

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Chemoenzymatic Synthesis of Sugar-Related Polyhydroxylated Compounds, Iminocyclitols and Their Derivatives As Glycosidase Inhibitors Chemoenzymatic synthesis of sugar-related polyhydroxylated compounds, iminocyclitols and their derivatives as glycosidase inhibitors Alda Lisa Concia ADVERTIMENT. La consulta d’aquesta tesi queda condicionada a l’acceptació de les següents condicions d'ús: La difusió d’aquesta tesi per mitjà del servei TDX (www.tdx.cat) i a través del Dipòsit Digital de la UB (diposit.ub.edu) ha estat autoritzada pels titulars dels drets de propietat intel·lectual únicament per a usos privats emmarcats en activitats d’investigació i docència. No s’autoritza la seva reproducció amb finalitats de lucre ni la seva difusió i posada a disposició des d’un lloc aliè al servei TDX ni al Dipòsit Digital de la UB. No s’autoritza la presentació del seu contingut en una finestra o marc aliè a TDX o al Dipòsit Digital de la UB (framing). Aquesta reserva de drets afecta tant al resum de presentació de la tesi com als seus continguts. En la utilització o cita de parts de la tesi és obligat indicar el nom de la persona autora. ADVERTENCIA. La consulta de esta tesis queda condicionada a la aceptación de las siguientes condiciones de uso: La difusión de esta tesis por medio del servicio TDR (www.tdx.cat) y a través del Repositorio Digital de la UB (diposit.ub.edu) ha sido autorizada por los titulares de los derechos de propiedad intelectual únicamente para usos privados enmarcados en actividades de investigación y docencia. No se autoriza su reproducción con finalidades de lucro ni su difusión y puesta a disposición desde un sitio ajeno al servicio TDR o al Repositorio Digital de la UB. No se autoriza la presentación de su contenido en una ventana o marco ajeno a TDR o al Repositorio Digital de la UB (framing). Esta reserva de derechos afecta tanto al resumen de presentación de la tesis como a sus contenidos. En la utilización o cita de partes de la tesis es obligado indicar el nombre de la persona autora. WARNING. On having consulted this thesis you’re accepting the following use conditions: Spreading this thesis by the TDX (www.tdx.cat) service and by the UB Digital Repository (diposit.ub.edu) has been authorized by the titular of the intellectual property rights only for private uses placed in investigation and teaching activities. Reproduction with lucrative aims is not authorized nor its spreading and availability from a site foreign to the TDX service or to the UB Digital Repository. Introducing its content in a window or frame foreign to the TDX service or to the UB Digital Repository is not authorized (framing). Those rights affect to the presentation summary of the thesis as well as to its contents. In the using or citation of parts of the thesis it’s obliged to indicate the name of the author. Chemoenzymatic synthesis of sugar-related polyhydroxylated compounds, iminocyclitols and their derivatives as glycosidase inhibitors Instituto de Química Avanzada de Cataluña (IQAC) Consejo Superior De Investigaciones Científicas (CSIC) Facultad de Química, Universidad de Barcelona, Departamento de Química Orgánica, Programa de Química Orgánica Alda Lisa Concia, 2012 CONSEJO SUPERIOR DE INVESTIGACIONES CIENTÍFICAS (CSIC) INSTITUTO DE QUÍMICA AVANZADA DE CATALUÑA (IQAC) UNIVERSIDAD DE BARCELONA Facultad de Química Departamento de Química Orgánica Programa de Doctorado de Química Orgánica Chemoenzymatic synthesis of sugar-related polyhydroxylated compounds, iminocyclitols and their derivatives as glycosidase inhibitors Memoria presentada por Alda Lisa Concia para optar al Título de Doctor por la Universidad de Barcelona Dirigida por: Tutor: Dr. Jesús Joglar Tamargo Dr. Francesc Rabanal Anglada Dr. Pere Clapés Saborit Doctoranda: Alda Lisa Concia A mio papà, per avermi insegnato l’importanza del sapere e avermi trasmesso la sua curiosità. AGRADECIMIENTOS El primer agradecimiento es para mis directores de tesis Jesús Joglar Tamargo y Pere Clapés Saborit. Antes que nada por la confianza depositada en mi persona y por permitirme formar parte de su grupo. Gracias por su gran disponibilidad, su asesoramiento y consejos en todo el proceso de investigación y por sus atinadas correcciones y sugerencias en la elaboración de la tesis. Gracias también por su comprensión y apoyo en los momentos críticos de mi vida personal. A mis compañeros de grupo, desde quien me acogió el primer día hasta quien ha acabado conmigo esta experiencia, quien ha sido una constante compañía y quien ha pasado cortas estancias, quien todavía está por aquí y quien se ha ido por otros lares. Gracias por ofrecerme generosamente sus conocimientos y experiencia, por alegrar las horas de trabajo en el laboratorio y por compartir conmigo unos geniales momentos: las comidas, los cafés (y cigarrillos), las cenas, las fiestas, despedidas, rural rave, island rave y mucho más. Gracias también y sobre todo, por la solidaridad y el afecto que me han brindado en algunos momentos difíciles. Un agradecimiento especial a Mariana con cuya ayuda siempre puedo contar desde el principio hasta el final y a Carles y Livia por su colaboración en este trabajo experimental y, por supuesto, por su apoyo y amistad. A todos mis verdaderos amigos, cercanos y lejanos, por haberme soportado con paciencia, haberme animado e impulsado en todo momento. Por ultimo, y no por ello menos importante, a mi madre y mi padre por las oportunidades que me han ofrecido, la comprensión con mi distancia y el amor con el cual me han acompañado en la experiencia de este doctorado. Gracias a todas estas personas porque afortunadamente en la vida, un ser humano si está solo, no es nada. 7 Summary Section 1 (Introduction) is a comprehensive review of the subjects discussed in this thesis: biocatalysis, aldolases and iminocyclitols. It contains a description of the application of dihydroxyacetone phosphate (DHAP) and dihydroxyacetone (DHA) utilizing aldolases to the chemoenzymatic synthesis of bioactive compounds and an introduction to the structure, biological activities and synthesis of iminocyclitols. Recent bibliographic references are included at the end of the section. Section 2 (Objectives) outlines the aims of this thesis. Section 3 (Results and discussion) describes the studies carried out in this thesis. Bibliographic references are included as an overview of previous results and to support our statements at the end of each chapter. - Section 3.1 deals with the application of D-fructose-6-phosphate aldolase in organic chemistry. It describes the chemoenzymatic preparation of polyhydroxylated compounds, sugars 1-deoxy-D-xylulose and 1-deoxy-D-ido-hept-2-ulose and iminocyclitols 1-deoxynojirimycin, 1-deoxymannojirimycin and N-alkylated derivatives, 1,4-dideoxy-1,4-imino-D-arabinitol and 1,4,5-trideoxy-1,4-imino-D-arabinitol. An unprecedented methodology, having as a key step a novel enzymatic aldol addition reactions catalyzed by D-fructose-6-phosphate aldolase, is presented. - Section 3.2 presents the cascade chemical-enzymatic synthesis of a collection of novel 1,4-dideoxy-1,4-imino-D- and -L-arabinitol (DAB and LAB) 2-aminomethyl derivatives including 2-oxopiperazine conjugates which have an interest as potential glycosidase inhibitors. - Section 3.3 describes the chemoenzymatic synthesis of novel polyhydroxylated pyrrolizidines of the family of casuarines by means of an asymmetric strategy based on cascade aldol additions catalyzed by dihydroxyacetone phosphate (DHAP) and dihydroxyacetone (DHA) aldolases. - Section 3.4 describes the methodology and results of the preliminary in vitro assays for glycosidases inhibition activity of some of the compounds synthetized during the course of this thesis. Section 4 (Experimental) describes the procedure for the experimental work carried out in this project. 1H and 13C NMR spectra can be found in the attached CD. 8 Contents 1. INTRODUCTION ............................................................................................................. 13 1.1. BIOCATALYSIS AND CHEMOENZYMATIC PROCESSES: GENERAL CONSIDERATIONS ....................... 15 1.2. ALDOLASES..................................................................................................................................... 16 1.2.1. Application of aldolases in organic synthesis .................................................................... 16 1.2.2. Classes of aldolases and their applications ....................................................................... 17 1.2.3. DHAP-dependent aldolases ............................................................................................... 22 1.2.4. DHA-dependent aldolases ................................................................................................. 27 1.3. IMINOCYCLITOLS ............................................................................................................................ 30 1.3.1. Definition and historical background ................................................................................ 30 1.3.2. Biological properties .......................................................................................................... 31 1.3.3. Therapeutic applications and mode of action ................................................................... 33 1.3.4. Polyhydroxylated pyrrolidines 1,4-dideoxy-1,4-imino-D- and -L-arabinitol……………………..41 1.3.5. Chemo-enzymatic synthesis of iminosugars and derivatives ............................................ 42 1.4. REFERENCES ................................................................................................................................... 46 2. OBJECTIVES ..................................................................................................................
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