University of Missouri, St. Louis IRL @ UMSL Dissertations UMSL Graduate Works 11-22-2019 Towards Completely Automated Glycan Synthesis Matteo Panza University of Missouri-St. Louis, [email protected] Follow this and additional works at: https://irl.umsl.edu/dissertation Part of the Organic Chemistry Commons, Polymer and Organic Materials Commons, and the Polymer Chemistry Commons Recommended Citation Panza, Matteo, "Towards Completely Automated Glycan Synthesis" (2019). Dissertations. 907. https://irl.umsl.edu/dissertation/907 This Dissertation is brought to you for free and open access by the UMSL Graduate Works at IRL @ UMSL. It has been accepted for inclusion in Dissertations by an authorized administrator of IRL @ UMSL. For more information, please contact [email protected]. Towards completely automated glycan synthesis By Matteo Panza Master of Science (Chemistry), University of Missouri-St. Louis, May 2017 Master of Science (Chemistry), University of Milan, Italy, October 2014 Bachelor of Science (Chemistry), University of Milan, Italy, October 2012 A Dissertation Submitted to the Graduate School of the UNIVERSITY OF MISSOURI – ST. LOUIS in Partial Fulfillment of the Requirements for the Degree of DOCTOR OF PHILOSOPHY in CHEMISTRY December 2019 Dissertation Committee Prof. Alexei V. Demchenko, Ph.D. (Chair) Prof. Eike Bauer, Ph.D. Prof. Keith J. Stine, Ph.D. Prof. Chung F. Wong, Ph.D. ABSTRACT Towards completely automated glycan synthesis Matteo Panza Doctor of Philosophy, University of Missouri – St. Louis Prof. Alexei V. Demchenko, Advisor Carbohydrates are ubiquitous both in nature as biologically active compounds and in medicine as pharmaceuticals. Although there has been continued interest in the synthesis of carbohydrates, chemical methods require specialized knowledge and hence remain cumbersome. The need for development of rapid, efficient and operationally simple procedures has come to the fore. This dissertation focuses on the development of a fully automated platform that will enable both experts and non-specialists to perform the synthesis of glycans. Existing automated methods for the synthesis of oligosaccharides are highly sophisticated, operationally complex, and require significant user know-how. By contrast, high performance liquid chromatography (HPLC) equipment-based automation introduced by our lab represents a highly accessible method of synthesis. This approach offers operational simplicity by delivering all reagents using standard liquid handling components and convenient real-time reaction monitoring of every step using detectors and standard computer software and interface. Many operations still require the operator intervention, and the entire technology remains semi-manual. Building upon promising preliminary results, this dissertation aimed to generate a universal platform for the fully automated synthesis of glycans. To achieve the complete automation of the solid phase synthesis that has the potential to revolutionize glycan synthesis, this dissertation focuses on the following aspects: 1) the development of new concepts for chemical glycosylation applicable to stereoselective formation of challenging 1,2-cis glycosidic bonds; 2) implementation of autosamplers and switch valves as new components to achieve operator-less automation; and 3) the development of new chemically stable resins for solid phase synthesis. With these key developments, we have acquired a reliable and simple platform for fully-automated oligosaccharide synthesis. The proof of concept was assessed by the synthesis of a number of target glycans. Synthesis of ii carbohydrates and other classes of biomolecules using this user-friendly and fully- automated platform will accelerate discovery in many scientific disciplines, most prominently chemistry, automation, and therapeutic-agents development. iii ACKNOWLEDGEMENTS I have a long list of acknowledgments and I would like to begin thanking Prof. Alexei Demchenko. In these years he has been more than an advisor, he has been a mentor and a friend, guiding me through science and life decisions. His passion and enthusiasm for science and his curiosity have set an example for my future career. He has always devoted his time to make me feel welcomed in his lab and to my research to be productive. I would like to thank my committee members, Prof. Eike Bauer, Prof. Keith J. Stine and Prof. Chung F. Wong for their encouragement and their patience all throughout my PhD. I am thankful to Dr. Jagodige P. Yasomanee for training me during my first year in Glycoworld. I would like to especially thank Dr. Rensheng Luo for the help with NMR and low temperature experiments. Also, I am very thankful to the funding received towards my PhD from the National Institute of Health (NIH) and University of Missouri- St. Louis. I would like to thank all faculty and staff from the Department of Chemistry and Biochemistry at UMSL for all the knowledge I have gained and support throughout my studies at UMSL. My thanks and affection will always be to my second family, my Glycoworld present and former lab members, Dr. Salvo Pistorio, Dr. Xiao Jia, Dr. Satsawat Visansirikul (First), Dr. Michael Mannino, Dr. Tinghua Wang, Dr. Mithila Bandara, Scott Geringer, Catherine Alex, Samira Escopy, Ganesh Shrestha, Melanie Shadrick, Dr Crystal O’Neil and Dr. Yashapal Singh for the good time together and for making me feel welcomed in St. Louis. Last, I am grateful to my wife, Myriam, for all the patience, the support, the encouragement and the motivation. Thanks to my daughter for always making my days plenty of fun. I want to thank my parents, my brother, my sister and my relatives for always believing in me and helping me in this American adventure. Thank you to my close friends Marta, Andrea, Manu, Nene and many others who have always been here for me and my family. iv LIST OF ABBREVIATIONS Å ............................................................................................................................Ångström Ac ...............................................................................................................................Acetyl Ac2O...........................................................................................................Acetic anhydride AcOH ..................................................................................................................Acetic acid AgOTf ..........................................................................Silver(I) trifluoromethanesulfonate Ag2CO3........................................................................................................Silver carbonate BF3•OEt2 ......................................................................................Boron trifluoride etherate BH3•THF...........................................................................Borane tetrahydrofuran complex Bn ...............................................................................................................................Benzyl BnBr ............................................................................................................Benzyl bromide Br2 ...........................................................................................................................Bromine Bz .............................................................................................................................Benzoyl BzCl............................................................................................................Benzoyl chloride BzCN...........................................................................................................Benzoyl cyanide CaH2 ...........................................................................................................Calcium hydride CBn................................................................................................................2-Cyanobenzyl CCl3CN............................................................................................... Trichloroacetonitrile CDCl3 ...............................................................................................Deuterated chloroform CD3COCD3............................................................................................ Deuterated acetone CH2Cl2 .......................................................................................................Dichloromethane CH3CN................................................................................................................Acetonitrile ClCH2CH2Cl ..........................................................................................1,2-Dichloroethane v Cu(OTf)2.....................................................................Copper(II) trifluoromethanesulfonate δ .....................................................................................................................Chemical shift d ................................................................................................................................Doublet DBU.............................................................................1,8-Diazabicyclo[5.4.0]undec-7-ene DFT................................................................................................Density functional theory dd ...........................................................................................................Doublet of doublets DIPEA.......................................................................................N,N-Diisopropylethylamine DMAP .........................................................................................4-Dimethylaminopyridine