Valencia, Tania, Yen, Laura Y., Berman, Cindy, Vincent, Thomas, Davis, Scott, Varrone, Francesca, Huang, Jianfeng, Mastroianni, Jessica, Carlson, Morgan, Owen, Tate, Kamel, Amin, Drygen, Denis, Kinberger, Garth, Gangwar, Shanti Pal, Yelshanskaya, Maria V, Ridley, John, Kirby, Robert, Alvarez, Jesus, Lakhia, Ronak, Patel, Vishal, Sobolevsky, Alexander I and Lee, Edmund C. (2025) The nucleobase guanine at the 3'-terminus of oligonucleotide RGLS4326 drives off-target AMPAR inhibition and CNS toxicity. Nature communications, 16 (1). p. 10762. ISSN 2041-1723

Abstract

Designing safe and effective oligonucleotide (ON) therapeutics requires thorough understanding of structural-activity relationship (SAR) with the intended on-target(s) as well as the unintended off-target(s). Despite encouraging pharmacodynamic activity in a Phase 1b study, development of the first-generation anti-miR-17 ON RGLS4326 for the treatment of autosomal dominant polycystic kidney disease was discontinued due to dose-limiting central nervous system (CNS)-related toxicity observed in nonclinical chronic toxicity studies. Here, we provide SAR evidence that the nucleobase guanine at the 3'-terminus of RGLS4326 drives an unexpected off-target aptamer-like direct interaction with α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid receptor (AMPAR), thereby causing CNS toxicity. By replacing the 3'-terminal guanine with adenine, we discover the next-generation anti-miR-17 RGLS8429 that is devoid of off-target AMPAR interaction and CNS toxicity while preserving the potency against the on-target miR-17. Here, we show a way to avoid off-target CNS effects and, more importantly, data that support the clinical development of RGLS8429.

Item Type:	Article
Keywords:	Guanine Humans Animals Oligonucleotides Receptors, AMPA MicroRNAs Central Nervous System Structure-Activity Relationship HEK293 Cells Mice
Date Deposited:	18 Dec 2025 00:45
Last Modified:	18 Dec 2025 00:45
URI:	https://oak.novartis.com/id/eprint/58142