Nucleic acid-templated assembly of therapeutic agents

Small molecule targeting of nucleic acids has come into focus as a therapeutic strategy for diseases such as myotonic dystrophy type 1 (DM1), a trinucleotide repeat disease characterized by its RNA gain-of-function mechanism. Methods of targeting nucleic acids range from the use of antisense oligonucleotides to the use of small molecules and oligomeric compounds to modulate symptoms. Of particular interest for targeting nucleic acid repeats is the use of multivalent targeting agents. Herein, we have explored the assembly of multivalent targeting agents using the nucleic acid target as a template for in situ synthesis. Using proximity driven azide-alkyne click, we developed a screening approach through which several hit compounds were found. As the proximity-induced click reaction is irreversible and requires long reaction times of at least 1 day before product is observed, we set out to develop a method for rapid, template-selected, reversible assembly of therapeutic agents in situ via aldehyde-amine condensation. After a proof-of-concept assay with compounds that contain two reactive aldehyde or amine groups, a library of rationally designed small molecule targeting agents containing aldehyde or amine functionality was synthesized and screened. The selective assembly of fragments on template was confirmed by MALDI-MS in the presence of DM1- and Huntington’s Disease (HD)-relevant nucleic acid sequences. Of interest for both DM1 and HD, the resulting hit combinations of aldehyde and amine inhibited the formation of toxic RNA in vitro in a cooperative manner with low micromolar IC50 values and rescued mis-splicing in DM1 model cells. This reversible template-selected assembly is a promising strategy to achieve multivalent targeting and could be utilized to develop strong and selective binders for other disease-relevant nucleic acid targets.

Name thesis:degree_name

Ph.D.

Level thesis:degree_level

Dissertation

Discipline thesis:degree_discipline

Chemistry

Grantor

University of Illinois at Urbana-Champaign

Year dc:date

2022

Author dc:creator

• Krueger, Sarah Bonson

Contributors dc:contributor

• Zimmerman, Steven C
• Mitchell, Douglas A
• Jin, Hong
• Olshansky, Lisa

• nucleic acid
• DM1
• transcription
• assembly
• dynamic covalent chemistry

Statement dc:rights

• Copyright 2022 Sarah Bonson Krueger

Language dc:language

en, eng

Handle dc:identifier

https://hdl.handle.net/2142/115687

OAI record oai:identifier

https://www.ideals.illinois.edu/oai-pmh?verb=GetRecord&metadataPrefix=etdms&identifier=oai%3Awww.ideals.illinois.edu%3A2142%2F115687