A Bioanalytical Liquid Chromatography Tandem Mass Spectrometry Approach for the Quantification of a Novel Antisense Oligonucleotide Designed for Parkinson's Disease: A Rat Brain Biodistribution Study.

IF 3.7 Q1 CHEMISTRY, MEDICINAL ACS Pharmacology and Translational Science Pub Date : 2025-02-05 eCollection Date: 2025-02-14 DOI:10.1021/acsptsci.4c00698

Anastasia Palaiologou, Marianna Naki, Marina Pantazopoulou, Fedon-Giasin Kattan, Leonidas Stefanis, Epaminondas Doxakis, Constantin Tamvakopoulos

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Abstract

Antisense oligonucleotides (ASOs) represent a unique category of therapeutics targeting disease-related RNAs. Since this new therapeutic category emerged, the immediate need to analyze ASOs in clinically relevant biological matrices has led to several methodologies, such as ligand binding assays and imaging techniques. To overcome issues in specificity and provide exact quantitative data for ASOs, a new LC-MS/MS method was developed to analyze in brain tissue a novel 4-10-4 gapmer ASO with the potential for treating Parkinson's disease with phosphorothioated backbone and 2'-O-(2-methoxyethyl) modifications. The sample pretreatment protocol to extract the ASO from brain tissue employed solid phase extraction (SPE) and protein digestion. The LC-MS/MS method was fully optimized, validated and applied to quantify the target ASO in brain tissue samples following an in vivo brain distribution study. The method has a Lower Limit Of Quantification of 1 ng/mg and was applied to the study's samples, demonstrating satisfactory sensitivity and providing valuable information about the ASO's distribution in different brain regions over 45 days.

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一种用于帕金森病的新型反义寡核苷酸定量的生物分析液相色谱串联质谱方法：大鼠大脑生物分布研究。

反义寡核苷酸（ASOs）是一类独特的靶向疾病相关rna的治疗药物。自从这一新的治疗类别出现以来，迫切需要分析临床相关生物基质中的ASOs，这导致了几种方法，如配体结合试验和成像技术。为了克服特异性问题并提供精确的定量数据，建立了一种新的LC-MS/MS方法来分析脑组织中具有磷酸化骨架和2'- o -（2-甲氧基乙基）修饰的新型4-10-4间隙分子ASO，该ASO具有治疗帕金森病的潜力。从脑组织中提取ASO的样品前处理方案采用固相萃取（SPE）和蛋白质消化。通过脑内分布研究，对LC-MS/MS方法进行了充分优化和验证，并将其应用于脑组织样品中目标ASO的定量。该方法的定量下限为1 ng/mg，应用于本研究的样品，显示出令人满意的灵敏度，并提供了有关ASO在45天内不同大脑区域分布的有价值的信息。

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来源期刊

ACS Pharmacology and Translational Science Medicine-Pharmacology (medical)

CiteScore

10.00

自引率

3.30%

发文量

133

期刊介绍： ACS Pharmacology & Translational Science publishes high quality, innovative, and impactful research across the broad spectrum of biological sciences, covering basic and molecular sciences through to translational preclinical studies. Clinical studies that address novel mechanisms of action, and methodological papers that provide innovation, and advance translation, will also be considered. We give priority to studies that fully integrate basic pharmacological and/or biochemical findings into physiological processes that have translational potential in a broad range of biomedical disciplines. Therefore, studies that employ a complementary blend of in vitro and in vivo systems are of particular interest to the journal. Nonetheless, all innovative and impactful research that has an articulated translational relevance will be considered. ACS Pharmacology & Translational Science does not publish research on biological extracts that have unknown concentration or unknown chemical composition. Authors are encouraged to use the pre-submission inquiry mechanism to ensure relevance and appropriateness of research.