Accessing Therapeutically-Relevant Multifunctional Antisense Oligonucleotide Conjugates Using Native Chemical...

Area of Science:

• Molecular Medicine
• Oligonucleotide Therapeutics
• Chemical Biology

Background:

• Antisense oligonucleotide (ASO) therapies offer precise gene expression modulation for various diseases.
• Challenges in intracellular targeting and delivery of ASOs necessitate advanced conjugation strategies.
• ASO-ligand conjugates are crucial for enhancing therapeutic efficacy and specificity.

Purpose of the Study:

• To develop a versatile method for conjugating ASOs with therapeutic ligands using native chemical ligation.
• To explore post-ligation functionalizations for creating multifunctional oligonucleotide constructs.
• To demonstrate the applicability of this methodology for clinically relevant ASO conjugates.

Main Methods:

• Utilized native chemical ligation to form stable amide linkages between ASOs and peptides/sugars.
• Incorporated therapeutically relevant modifications like locked nucleic acids and phosphorothioate backbones.
• Performed post-ligation functionalizations including radical desulfurization, lipidation, and alkylation.
• Applied the method to create a triantennary-GalNAc ASO conjugate.

Main Results:

• Successfully conjugated ASOs with various ligands and chemical modifications.
• Demonstrated a range of post-ligation functionalizations yielding diverse molecular handles (alkyne, biotin, chelators).
• Validated the binding and functional activity of a GalNAc-ASO conjugate, confirming applicability.

Conclusions:

• Native chemical ligation provides a robust platform for creating functionalized ASO-ligand conjugates.
• The developed methodology enhances ASO delivery and therapeutic potential.
• This approach is applicable to the development of next-generation oligonucleotide-based therapeutics.