Click chemistry for enhanced and emerging biological applications: from novel dyes for cell proliferation assays to mRNA-based vaccination

Montiel Campanón, Luis (2023): Click chemistry for enhanced and emerging biological applications: from novel dyes for cell proliferation assays to mRNA-based vaccination. Dissertation, LMU München: Fakultät für Chemie und Pharmazie

Abstract

The present dissertation represents a focused endeavor aimed at broadening click-chemistry-based applications in the field of nucleic acids. The principal objective is to extend the synthetic utility of bioorthogonal click chemistry by delving into the following diverse, yet equally crucial areas of research: DNA fluorescent-labeling for bioimaging purposes and ligand development for nucleic acid cell targeting in emerging therapeutics. We aimed to develop new fluorescent probes to improve detection sensitivity and quality of imaging methods. To achieve this, we established a divergent synthetic pathway that produced ready-to-click, pH insensitive rhodamine dyes with outstanding brightness, which were proven to be a superior alternative in EdU cell proliferation assays. Furthermore, we prepared a group of fluorescein- and rhodamine-based multivalent dendrons by branching click chemistry, which exhibited self-quenching effects and demonstrated potential application as FRET quenchers. Additionally, in an effort to develop emerging therapeutics using click chemistry, particularly in vaccination and cancer immunotherapies, we synthesized three distinct ready-to-click mannose functionalized ligands with the aim of targeting dendritic cells (DCs). Our analyses confirmed the uptake of the ligands’ fluorescent conjugates into DCs through endocytosis. Finally, spurred by the COVID-19 pandemic, baseclick GmbH took on the task of developing a SARS-CoV-2 vaccine candidate. To this aim, we utilized our DC targeting ligands and baseclick’s mRNA labeling technology to successfully synthesize a mannose mRNA conjugate. At present, pharmacokinetics and pharmacodynamics of the vaccine are under investigation. The promising in vitro results, nevertheless, instills optimism in our efforts to establish a framework for future exploration of cell targeted, click-chemistry-based nucleic acid therapies.

• Dokumententyp: Dissertationen (Dissertation, LMU München)
• Themengebiete: 500 Naturwissenschaften und Mathematik ; 500 Naturwissenschaften und Mathematik > 540 Chemie
• Fakultäten: Fakultät für Chemie und Pharmazie
• Sprache der Hochschulschrift: Englisch
• Datum der mündlichen Prüfung: 19. September 2023
• 1. Bericht­erstatter:in: Carell, Thomas