Challenges in the Stability, Safety, and Efficacy of Antisense Oligonucleotide Therapeutics

Abstract: Antisense oligonucleotides represent a promising class of therapeutic agents due to their high specificity in modulating gene expression. These short, single-stranded synthetic sequences (typically 13–25 nucleotides) function by binding to complementary RNA targets, thereby influencing RNA processing or translation, primarily through RNase H-mediated degradation. Over the past decade, significant advances in chemical modifications and delivery strategies have led to a notable increase in the number of ASO-based therapies that have reached the market. Despite this progress, several challenges persist, including poor stability in biological fluids, limited cellular uptake, and safety concerns related to off-target effects and immunogenicity. This review provides a comprehensive overview of the mechanisms of action of ASOs, their pharmacological applications, current market status, and clinical progress. Special attention is given to evaluating in vitro and in vivo ASO stability using nanotechnology-based delivery approaches to improve pharmacokinetics/ pharmacodynamics and structural modifications to enhance therapeutic potential. Additionally, we discuss the associated adverse effects and propose strategies to mitigate them.