Short review on determination of nusinersen for spinal muscular atrophy treatment

Abstract

Nusinersen, an antisense oligonucleotide, is an active ingredient of the first drug approved by the Food and Drug Administration for the treatment of spinal muscular atrophy, a genetic condition characterized by progressive muscle weakness and atrophy. A deficiency in the survival motor neuron (SMN) protein causes the disease. SMN protein is required for the functioning of motor neurons and the transmission of nerve signals to muscles. Using nusinersen therapy determines the need for a pharmacokinetic and metabolic analysis of the drug. Consequently, this requires the use of various analytical techniques to study biological samples. Based on current scientific literature, encompassing publications from 2007 to 2024, the sample preparation methods for nusinersen involve liquid-liquid extraction, solid-phase extraction, dispersive solid-phase extraction, and microextraction by packed sorbent or hybridization, which facilitate the removal of interfering components and ensure the efficient recovery of the analyte. Liquid chromatography combined with mass spectrometry is currently the primary tool used for the determination of nusinersen and its metabolites. This review discusses the latest developments in analytical methods used in nusinersen research, presents techniques for the purification of biological samples, and compares their efficiency. The use of ultra-performance liquid chromatography coupled with mass spectrometry for the determination of oligonucleotides is also described. The results obtained using different chromatographic modes (reversed-phase high-performance liquid chromatography, ion-pair reversed-phase high-performance liquid chromatography, and hydrophilic interaction liquid chromatography) are then summarised and compared.